Recovery and Lifespan of Red Blood Cells From Pathogen-reduced, Stored Blood Units
Recovery and Lifespan of Red Blood Cells From Pathogen-reduced, Stored Blood Units
The pilot study has two objectives: 1) to assess the post-infusion viability of INTERCEPT RBC
 by measuring the 24 hour post-infusion recovery ("PTR24") and lifespan of autologous RBCs
 prepared with the INTERCEPT System for RBC after storage for 35 days under standard blood
 banking conditions using two different RBC labels; 51-chromium and biotin. The control will
 be conventional untreated RBCs stored for 35 days; and 2) comparison and contrast of PTR24
 and lifespan results of the 51-chromium and biotin labeling methods of RBC stored for 35 days
 under standard blood banking conditions. The purpose of gathering these data is to obtain
 more meaningful survival data for stored conventional and INTERCEPT RBCs over the entire 120
 d RBC lifespan (51-Cr labeled RBC permits a maximum 28 d assessment as a result of 51-Cr's
 variable, progressive elution from RBC and radioactive half-life).
Each subject will receive one infusion of autologous, radiolabeled and BioRBC labeled
 INTERCEPT RBCs (Test RBCs) and one infusion of autologous BioRBC labeled untreated RBCs
 (Control RBCs) concomitantly. Each infusion will be approximately 20 mL, i.e., 10 mL of
 51-chromium labeled RBC, and 10 mL of either BioRBC-6 or BioRBC-18 which will be stratified
 as indicated above based on the Test or Control designation.

 Treatment Compliance Healthy subjects who understand the study commitments and sign the
 informed consent will be enrolled in the study. Treatment compliance and follow-up testing
 will be tracked by the Investigator, recorded on the case report form (CRF), and monitored by
 the Sponsor.

 ASSESSMENT OF EFFICACY

 Efficacy Parameters

 Efficacy endpoints include the following:

 Primary Efficacy Endpoint(24)

 - 24 hour post-infusion recovery of autologous RBC stored in AS-5 for 35 days and then
 labeled with either 51Cr or biotin.

 Secondary Efficacy Endpoints(25)

 -  Mean lifespan of autologous RBC stored for 35 days and then labeled with either 51Cr or
 biotin

 -  Median lifespan (T50) of autologous RBC stored for 35 days and then labeled with either
 51Cr or biotin

 -  Area under the curve (AUC) determined using data points collected forRBC lifespan of
 autologous, radiolabeled or BioRBCs

 e) Dose determination and duration

 Methods and Timing of Efficacy Parameters This study consists of a single treatment period
 where Control biotinylated (using a RBC surface density of 6 or 18 mcg of sulfo-NHS-biotin
 per mL RBC) are administered, i.e., co-infused at a single point in time with Test
 INTERCEPT-treated biotinylated RBC (using a concentration of 18 or 6 mcg of sulfo-NHS-biotin
 per mL RBC) and INTERCEPT-treated, radiolabeled (51Cr) RBC. All 6 subjects will receive both
 control and S-303-treated (test) RBCs concomitantly and the only difference between the two
 sets of 3 subjects will be the level of biotinylation of the control or test RBC.

 The study is stratified and control or test RBC will be labeled alternatively with either 6
 or 18 mcg of sulfo-NHS-biotin per mL RBC. RBC from three individuals (subjects 1, 3 and 5)
 will be labeled with 6 and 18 mcg of sulfo-NHS-biotin per mL RBC for control and test RBC,
 respectively. Other three individuals (subjects 2, 4 and 6) will be labeled with 6 and 18 mcg
 of sulfo-NHS-biotin per mL RBC for Control and Test RBC, respectively. The objective of this
 stratification is to ensure that no bias is introduced by the biotin label density into one
 of the arms of the study.

 f) Description of observations and measurements

 On Day 0 of each treatment period the subject will donate two units of packed RBC (double RBC
 automatized donation, Trima - Terumo BCT apheresis system); the units will be leukocyte
 reduced by in-line filtration and the RBC concentrates will be prepared in additive solution
 (AS-5).

 One of the two units will be stored immediately at 1-6ºC for 35 days (Control unit). This
 conventional RBC component represents the Control RBCs in this study.

 The Test RBC unit will be prepared by processing using the INTERCEPT Blood System for Red
 Blood Cells. In vitro biochemical and morphological parameters of the study RBC components
 (Table 1) will be measured on Day 0 after preparation of RBC concentrates in AS-5 (prior to
 study treatment), post processing and prior to storage (Test only), and again on Day 35
 (prior to infusion back in to study subjects) to evaluate the RBC quality of the
 processed/stored RBC units. The data collected for the in vitro characteristics will be
 summarized using descriptive statistics.

 Table 1. Biochemical parameters analyzed on day 0 and day 35 of storage of control- and
 treated RBC units.

 Parameter Hb content/unit Hemolysis pH pO2 pCO2 Glucose (extracellular) Lactate
 (extracellular) Bicarbonate (extracellular) Sodium (extracellular) Potassium (extracellular)

 On Day 35 of RBC storage each treatment period, an aliquot of Control RBCs will be
 biotinylated (6 or 18 mcg/mL). From the Test unit, two aliquots will be drawn; the first
 aliquot will be radiolabeled with 51Cr and the second aliquot will be biotinylated at 6 or 18
 mcg/mL. Re-infusion of the two biotinylated autologous RBC (Control and Test) specimens along
 with the 51Cr labeled autologous RBC (Test) will measure recovery and survival (approximately
 total: 20 mcCi). Also on Day 35, a 20 mL sample of fresh blood will be drawn from the subject
 and the RBCs labeled with 99mTc and Bio-54. Re-infusion of the fresh 99mTc RBC and fresh
 Bio-54 will allow determination of subject blood volume or number of circulating RBC.

 Five types of RBC will be mixed and infused: a) fresh RBC labeled with 99mTc and BioRBC-54;
 b) fresh Bio-54 labeled RBC; c) Stored, Control or Test RBC biotinylated at density 6 mcg/mL;
 d) Stored, Test or Control RBC biotinylated at density 18 mcg/mL; and, e) Stored, Test, RBC
 labeled with 51Cr. All aliquots will be labeled separately and mixed for a volume of ~50 mL.
 The 50 mL of autologous RBCs will be infused through a peripheral vein using an 18-19 gauge
 butterfly needle (at an approximate rate of 5 mL/min).

 Blood samples, from the subject's contralateral arm, will be collected immediately
 pre-infusion and at 5, 7.5, 10, 12.5, 15, 20, 30 minutes and 24 hours after completion of the
 infusion to permit extrapolation to T=0 for 51Cr determination of blood volume as previously
 published by our group (16, 17, 26-30).

 Additional blood samples will be collected at 48 and 72 hours and 7 days post-infusion, and
 then weekly through 28 days (days 37, 38, 42, 49, 56 and 63 post-infusion) and bi-weekly from
 then on (days 77, 91, 105, 119, 133, 147). After collection of the final blood sample in
 Treatment Period 2, subjects will be discharged from the study.

 Upon completion of the study, each of the 6 subjects will have provided 2 units of whole
 blood and receive 1 infusion of autologous radiolabeled Test or Control RBCs and 1 infusion
 of autologous radiolabeled Control or Test RBCs.

 The study assessments used to evaluate both efficacy and safety are presented below (Section
 1).

 g) Clinical procedures, laboratory tests, monitoring drug effects and minimizing risks

 Section 1: Study Assessments and data collection Screening: Day -28 to 0

 -  Explain study to subject

 -  Obtain signed Informed Consent

 -  Medical history

 -  Blood donor physical exam

 -  Vital signs

 -  Concomitant medications, i.e., other than those excluded

 -  Samples for laboratory tests: Hematology, serum Chemistry, and Blood Donor testing
 (microbiological, ABO/Rh) screening panel (following FDA/AABB requirements).

 -  Direct and Indirect Antiglobulin Test (DAT/IAT)

 -  ABO and Rh type (part of the donor panel)

 -  Serum or plasma sample to test for antibody specific to INTERCEPT RBC and antibody
 specific to biotinylated RBC

 -  Stratify subjects to receive BioRBC-6 or BioRBC-18 for INTERCEPT treated RBCs

 -  Serum or plasma sample for iron status testing: ferritin (being acceptable, ferritin >
 26 ng/mL) Prior to Blood Collection, Day 0

 -  Intercurrent illness

 -  Vital signs

 -  Concomitant medications

 -  Hemoglobin/hematocrit

 Double RBC Collection, Day 0 Trima Collection Procedure Parameters and Schedule (AC Ratio,
 etc.)

 1. Anticoagulant (AC) ratio The AC ratio is a parameter that is configurable on the Trima
 System. The red cells units will be collected at a whole blood to anticoagulant ratio of
 11.

 2. Run time The run time will be limited to a maximum of 120 minutes. Double RBC products
 will be collected when using the Trima RBC, Plasma sets and are expected to take
 approximately 30 minutes total.

 RBC Component - Prior to Study Treatment Day 0

 - Collect sample from the study Control and Test RBC units for in vitro lab assessments

 RBC Component - Study Treatment, Day 0-1

 -  Process and/or perform INTERCEPT Process for RBCs in one of the two donated blood
 units.

 -  Collect a sample from the Test RBC component (in vitro assessments)

 -  Place Control and Test RBC components in traditional blood bank storage (1-6ºC).

 Post Blood Collection, Day 1 (24±6 hours post donation)

 -  Concomitant medications

 -  Intercurrent illness

 -  Collect AEs and SAEs (Note: These data may be collected by phone)

 RADIOLABELING, BIOTINYLATION AND RE-INFUSION OF AUTOLOGOUS RBC ALIQUOTS Prior to
 Re-Infusion of autologous labeled, Day 35 (within 6 hours prior to infusion)

 -  Collect a fresh 20 mL sample of subject's blood for 99mTc and BioRBC-54 labeling

 -  ABO type

 -  Confirm the identity of the RBC study component by clerical analysis of
 chain-of-custody documentation and blood bag labels following industry standards
 and AABB/FDA regulations

 -  Samples for laboratory tests: Hematology and serum chemistry

 -  Pregnancy test (if female subject)

 -  Serum or plasma samples for INTERCEPT antibodies RBC and anti-BioRBC antibodies

 -  Samples for RBC recovery, mass, lifespan

 -  Intercurrent illness

 -  Vital signs

 -  Height and weight

 -  Concomitant medications

 -  Collect AEs and SAEs since donation.

 RBC Component:

 Prior to Infusion, Day 35

 - Label 10 mL aliquot of 35 day-old study RBC with 51Cr

 -  Label fresh 10 mL aliquot of subject's RBC with 99mTc and BioRBC-54

 -  Collect a sample of study RBC component (for in vitro assessments)

 Infusion of Autologous Study RBCs, Day 35

 - Infusion of a mixture of five ~10 mL (total ~50 mL) aliquots of autologous blood (all
 mixed together and infused at rate of 5-7 mL/min) including: a) fresh RBC labeled with
 99mTc and BioRBC-54; b) fresh Bio-54 labeled RBC; c) Stored, Control or Test RBC
 biotinylated at density 6 mcg/mL; d) Stored, Test or Control RBC biotinylated at density
 18 mcg/mL; and, e) Stored, Test, RBC labeled with 51Cr. All aliquots will be labeled
 separately and mixed for a volume of ~50 mL. The 50 mL of autologous RBCs will be
 infused through a peripheral vein using an 18-19 gauge butterfly needle (at an
 approximate rate of 5 mL/min). (Note: The remainder of the two 35 day-old INTERCEPT
 treated and non-treated Control RBC units are discarded.)

 -  Concomitant medications

 -  Collect AEs and SAEs

 -  Hematology tests: CBC including hemoglobin, hematocrit, platelets, MCV, MCH,
 MCHC, RDW ** Serum chemistry tests: calcium, bicarbonate, chloride, inorganic
 phosphate, potassium, sodium, cholesterol, glucose, total protein,
 triglycerides, LDH, ALT, AST, total bilirubin, BUN, & creatinine.

 -  Includes: serum or plasma ferritin, iron, iron binding capacity,
 transferrin saturation † Vital signs: heart rate, respiratory rate, blood
 pressure and temperature. If fever within 24 hours of infusion
 (temperature>39°C or >38˚C with shaking chills) culture the subject and
 the RBC bag.

 The assessments and data collection beginning the day following the RBC infusion until
 Study Day 147 (day 112 post-infusion) are shown in the Table 1.

 ASSESSMENT OF SAFETY The timing of study safety assessments is presented in the Table.

 Safety Parameters

 Safety endpoints include the following:

 Primary Safety Endpoint

 -  Incidence of antibody specific to INTERCEPT treated RBCs

 -  Incidence of antibody specific to biotinylated RBCs Secondary Safety Endpoint

 -  Incidence of adverse and severe adverse events as described below in the section
 titled, "Recording and Reporting Adverse Events." The INTERCEPT Treatment System
 medical device will be monitored for device deficiencies, including malfunctions
 and adverse device effects.

 Methods and Timing of Safety Parameters In this study, the Sponsor/Investigator, J.A.
 Cancelas, will collect and record adverse events (AEs) for 24 hours following collection
 of each whole blood unit and for 24 hours following each infusion of study RBCs. Serious
 adverse events (SAE) will be collected and recorded from the time the first unit of
 whole blood is collected until the completion of the study (Table).

 Subjects will be actively monitored for AEs during blood collection and during the study
 RBC infusions, and until they are discharged. After discharge, all AEs reported to the
 qualified study staff will be recorded. AE and SAE data may be collected by phone or in
 person by study staff. Vital signs are collected prior to and following each study
 infusion. In the event that a subject develops within 24 hours after a study RBC
 transfusion post-infusion fever, defined as a temperature >39°C or a temperature >38°C
 with shaking chills (rigors) or signs and symptoms consistent with bacterial sepsis, the
 subject's blood will be cultured and the Blood Center will be notified to culture the
 RBCs remaining in a segment or the residual Test or Control RBC components.

 Our group will test subject serum or plasma for antibody specific to INTERCEPT RBCs and
 BioRBC; these samples will be obtained from each subject pre-infusion, 14 days after
 infusion, and 28 days after infusion and monthly thereafter to study day 147 (see
 Table). If an antibody specific to INTERCEPT treated RBCs is identified, samples will be
 sent to an independent laboratory for confirmation (the UCLA/American Red Cross,
 Pomona). Blood samples for hematology and chemistry panels will be obtained from each
 subject Study Days: pre-infusion (day 35), 24 hours after the infusion (day 36), 56 days
 after infusion (day 91) and 112 days after infusion (day 147, at the end of the study).
 Clinically significant laboratory findings will be recorded as adverse events. The
 laboratory assessments are identified in the Table 1.

 Transfusion Reactions Although transfusion reactions are unlikely due to the small
 infusion volume, clinically significant transfusion reactions will be recorded as
 adverse events.

 Suspected Transfusion Related Sepsis Subjects will be monitored for infusion-related
 adverse events including infusion-related sepsis following the study infusion. To
 provide an unaltered sample of the blood infusate aliquot transfused into a subject, the
 remainder of this aliquot will be sent for bacterial culture.

 In the event that a subject develops post-infusion fever, defined as a temperature >39°C
 or a temperature >38°C with shaking chills (rigors) or signs and symptoms consistent
 with bacterial sepsis within 24 hours after a study RBC infusion, the subject's blood
 will be cultured and the remaining stored component will be cultured.

 Cultures will be performed by investigators at the Hoxworth Blood Center according to
 standard operating procedures at the Hoxworth Blood Center. If bacteria are recovered
 from both the patient's blood and the RBC component, samples of the isolates will be
 sent to the University of Cincinnati Hospital laboratory to determine if the recovered
 organisms are identical. Infusion-induced sepsis will not be established unless the same
 organism(s) is recovered from both the RBC component and from the study subject.

 Detection and Confirmation of Antibody Specific to INTERCEPT RBCs Although not
 anticipated to occur in this study due to the limited exposure to Test RBC, all subjects
 will be tested for the presence of antibody specific to INTERCEPT treated RBC at study
 entry (naturally occurring antibody) and at specified times during the study (Table 1).

 The presence of antibody will be evaluated using a formally validated gel card IAT that
 is routinely used in many blood centers for transfusion compatibility testing in
 conjunction with a panel of frozen reagent INTERCEPT RBCs. The frozen RBC screening
 panel is composed of reagent RBCs from 3 specifically phenotyped blood group O donors,
 and is analogous to routine antibody screening panels used in regular blood banking
 practice. Each cell in the panel of 3 consists of untreated RBCs, cells carrying a
 "high" level of acridine adducts and cells carrying a "low" level of acridine adducts.
 The screening panel was designed as a sensitive screening test for the presence of
 INTERCEPT RBC antibodies in the presence of most common alloantibodies. Gel card
 agglutination will be scored 1+ to 4+ based on manufacturer's recommendation.

 For result interpretation, if the RBC screening panel score is positive with 3/3 of the
 INTERCEPT RBCs carrying either 'high" and/or "low" levels of acridine adducts AND with
 0/3 with the corresponding Control untreated RBCs the sample is classified as "reactive
 for INTERCEPT RBC antibodies.

 Antibody confirmation and characterization will be performed at the ARC Immunohematology
 Reference Laboratory, Pomona and will include acridine specificity by agglutination
 inhibition (neutralization) using a soluble acridine analog of amustaline, antibody
 titer, antibody isotype and thermal amplitude.

 If the presence of antibody specific to INTERCEPT RBC is established, this does not
 determine whether that antibody is physiologically active or clinically significant,
 this can only be determined by any one of the following criteria indicative of
 accelerated RBC clearance in the absence of active bleeding, organ-mediated RBC
 sequestration, severe erythroid hypoplasia or other concurrent medical cause for acute
 anemia not associated with transfusion.

 Detection and Confirmation of Antibody Specific to BioRBC As with INTERCEPT RBCs,
 subjects will be screened prior to transfusion for the presence of naturally occurring
 or pre-existing antibody to BioRBC (18). This will be done using the same gel card
 agglutination as for the INTERCEPT RBCs but using BioRBCs incubated with subject serum
 or plasma. Briefly, 50 µL of 0.8% hematocrit target BioRBC-N or unlabeled RBC will be
 prepared from a blood GpO donor and added to the gel column. Subject plasma (25 µL) is
 added to the target RBCs and incubated at 37˚C for 30 min. Gel cards are subsequently
 centrifuged for 10 min. The presence of BioRBC agglutination is quantified using the
 manufacturer's gradation-specific definitions of reactivity. Reactivity scores will be
 based on manufacturers recommendation 0-4+ but, in addition, a score of 1+ (i.e.,
 agglutination in lower half of matrix) will be further sub-classified into three
 increasingly reactive 1+ designations: 1) ± indicates a slanted pellet with a few
 agglutinated RBC; 2) 1w indicates a slanted pellet with more agglutinated RBC; and 3) 1+
 indicates a disrupted RBC pellet with more agglutinated cells. As with the detection of
 INTERCEPT-specific antibodies, the detection of specific antibodies to BioRBC does not
 determine whether that these antibodies are physiologically active or clinically
 significant.

 Confirmatory testing of the induction of BioRBC-induced antibodies will be performed by
 agglutination inhibition (neutralization) with biotin compounds, e.g., biocytin,
 biotinylated gelatin or biotinylated albumin is able, are able to neutralize anti-BioRBC
 antibodies. So far, all identified preexisting, naturally occurring anti-BioRBC
 antibodies are neutralized by biotin compounds, while the induced anti-BioRBC antibodies
 are expected to be inhibited by BioRBC and not by free biotin as previously published by
 our group (5).

 Recording and Reporting Adverse Events Adverse Event Definition An AE is defined as any
 untoward medical occurrence in a patient or clinical investigation subject administered
 a pharmaceutical product and which does not necessarily have a causal relationship with
 this treatment. An adverse event can be any unfavorable and unintended sign (including
 an abnormal laboratory finding), symptom, or disease temporally associated with the
 medicinal product (in this study BioRBC and INTERCEPT RBC) whether or not related to the
 medicinal product (International Conference on Harmonization of Technical Requirements
 for Registration of Pharmaceuticals for Human Use (ICH-E2A, section 2.1)

 Serious Adverse Event Definition

 Some AEs are considered "serious" when the outcome of the event poses a threat to a
 patient's life or functional status. A serious adverse event (SAE) is an AE that results
 in the following outcomes:

 -  Results in death

 -  Life-threatening

 -  Requires inpatient hospitalization or prolongation of existing hospitalization

 -  Results in persistent or significant disability/incapacity

 -  Is a congenital anomaly/birth defect

 -  Important medical events that may not result in death, be life-threatening, or
 require hospitalization may be considered serious when, based upon appropriate
 medical judgment, they may jeopardize the subject and/or may require intervention
 to prevent one of the outcomes listed in this definition (ICH E2A section II/B).

 Adverse Device Effect Definition An Adverse Device Effect (ADE) is defined as an adverse
 event related to the use of an investigation medical device.

 Unanticipated Adverse Effect Definition An unanticipated adverse effect (UAE) is defined
 as any serious adverse effect on health or safety or any life-threatening problem or
 death caused by, or associated with, the process, if that effect, problem, or death was
 not previously identified in nature, severity, or degree of incidence in the
 investigational plan or application (including a supplementary plan or application), or
 any other unanticipated serious problem associated with a device that relates to the
 rights, safety, or welfare of subjects (21 CFR 812.3).

 Reporting Adverse Events AEs will be documented on the appropriate case report form
 (CRF), regardless of whether or not they are classified as associated with the study
 product.

 If an AE meets the criteria of a serious adverse event (SAE) or unanticipated adverse
 device effect (UAE) as described above, the Sponsor/PI will halt the study and notify
 the FDA within 7 days.

 The Investigators will comply with the applicable regulatory requirement(s) related to
 the reporting of serious adverse events to the IRB. An Investigator shall submit to the
 Sponsor and to the reviewing IRB a report of any unanticipated adverse device effect
 occurring during an investigation as soon as possible, but in no event later than 10
 working days after the Investigator first learns of the effect.

 The Sponsor will report suspected unexpected serious adverse reactions and/or
 unanticipated adverse device effects to comply with the applicable regulatory
 requirement(s).

 Assessing Seriousness and Severity of an Adverse Event The Investigator is required to
 assess if the adverse event is non-serious or serious, the latter is defined as an event
 that resulted in any of the following outcomes: death, a life-threatening event (results
 in an immediate risk of death from the reaction as it occurred), inpatient
 hospitalization or prolongation of existing hospitalization, a persistent or significant
 disability/incapacity, a congenital anomaly/birth defect, or another important medical
 event.

 The term "severe" is used to describe the intensity (severity) of a specific event (as
 in mild, moderate, or severe); the event itself, however, may be of relatively minor
 medical significance (such as severe headache). Severity is not the same as "serious",
 which is based on patient/event outcome or treatment criteria associated with events
 that pose a threat to a patient's life or functional status.

 Seriousness (not severity) serves as a guide for defining regulatory reporting
 obligations. The National Cancer Institute Common Terminology Criteria for Adverse
 Events (CTCAE, version 4.03) is used to assess and classify the intensity/severity of an
 event.

 Assessing Causal Relationship of an Adverse Events The Investigator will assess whether
 there is a reasonable possibility that the study blood collection or RBC infusion caused
 or contributed to an adverse event.

 Determination of whether there is a reasonable possibility that the study RBC component
 caused or contributed to an adverse event includes assessing temporal relationships,
 biologic plausibility, dechallenge/rechallenge information (if available), association
 (or lack of association) with underlying disease, and presence (or absence) of a more
 likely cause.

 The scale for assessing the causal relationship of an adverse event to the study RBC
 component (imputability levels) follows:

 Causality Assessment Scale Imputability Level Explanation Excluded When there is
 conclusive evidence beyond reasonable doubt for attributing the adverse event to
 alternative causes. Unlikely When the evidence is clearly in favor of attributing the
 adverse event to causes other than the study RBC component.

 1. Possible: When the evidence is indeterminate for attributing adverse event either
 to the RBC component or to alternative causes.

 2. Likely: Probable When the evidence is clearly in favor of attributing the adverse
 event to the RBC component.

 3. Certain: When there is conclusive evidence beyond reasonable doubt for attributing
 the adverse event to the RBC component.

 Assessing Intensity of an Adverse Events The Investigator is required to assess the
 intensity (severity) of each adverse event using the National Cancer Institute Common
 Terminology Criteria for Adverse Events (CTCAE)

 Adverse Event Follow up All SAEs will be followed until resolution, or the Investigator
 judges the event to be chronic or stable.

 Stopping Rules

 The following SR will be applied to all study subjects throughout the trial whenever
 this study's sponsor/PI (José Cancelas, M.D., Ph.D.,) and/or Clinical Monitor (Patricia
 Carey MD, Professor of Pathology, Medical Director, Hoxworth Blood Center)) agree that
 any of the following have occurred:

 1. An SAE that is determined to be possibly, probably, or definitely related to the
 infusion of autologous BioRBC;

 2. An AE related to the infusion of autologous BioRBC jeopardizes the subject's health
 or safety;

 3. An AE requiring medical or surgical intervention related to the infusion of
 autologous BioRBC is required to prevent occurrence of an SAE;

 4. A post-infusion of hemolysis event as assessed by anti-BioRBC antibody detection
 using the IgG gel card assay developed by our group.

 If any of the above conditions occur, the study will be paused and reported to FDA and
 IRB as required by current regulations to assess continuation of the study. Infused
 subjects will remain being followed as long as needed to identify the consequences of
 the AEs identified. Since there are no additional experimental BioRBC transfusions
 administered as part of this study, "continuation of the study" is understood to mean a
 continuation of the monitoring of biologic samples as already outlined in this study,
 but possibly including other testing as deemed appropriate by the study's sponsor/PI,
 Clinical Monitor, FDA and/or IRB.
Subject Inclusion Criteria

 -  Up to 12 subjects will be enrolled for a maximum of 6 eligible subjects in the pilot
 study to provide preliminary RBC survival data to support a subsequent adequately
 powered study capable of addressing the two objectives as stated above.

 -  Age ≥18 years, of either gender

 -  Normal health status (as determined by the Investigator review of medical history and
 blood donor physical exam)

 -  Weight over 140 lbs.

 -  Complete blood count (CBC; including RBC indices MCV, MCH, MCHC, and RDW) and serum
 chemistry values within normal limits (including calcium, bicarbonate, chloride,
 inorganic phosphate, potassium, sodium, cholesterol, glucose, total protein,
 triglycerides, LDH, ALT, AST,total bilirubin, BUN, creatinine). Values outside of
 normal reference range if considered not to be clinically significant may be allowed
 with a protocol exception.

 -  Minimum hemoglobin levels of 13 g/dL for female and 14 g/dL for male subjects

 -  Negative blood donor screening test panel for HIV, HBV, HCV, HTLV, Syphilis, WNV and
 Zika virus

 -  Female subjects of childbearing potential and male subjects must agree to use a
 medically acceptable method of contraception throughout the study periods. A barrier
 method of contraception must be included, regardless of other methods.

 -  Meet or exceed AABB guidelines for blood donation (with the exception of travel
 deferrals).

 -  Signed and dated informed consent form

 Subject Exclusion Criteria

 -  Clinically significant acute or chronic disease (as determined by the Investigator)

 -  History of RBC autoantibodies/autoimmune hemolytic anemia, RBC alloantibodies or
 autoimmune disease

 -  History of congenital red cell disorders including glucose-6-phosphate dehydrogenase
 (G6PD) deficiency

 -  Positive Direct (DAT) and Indirect antiglobulin test (IAT)at study entry

 -  Immunosuppressive therapy (e.g., oral or IV prednisone) within the past 28 days

 -  Treatment with any medication known to affect RBC viability

 -  Pregnant or nursing female

 -  Male subjects or female subjects of childbearing potential not using effective
 contraception

 -  Participation in another clinical study currently or within the past 28 days

 -  Prior exposure to INTERCEPT treated or BioRBCs

 -  Pre-existing antibody specific to INTERCEPT RBCs or BioRBC

 -  Subjects are excluded if receiving immunosuppressive therapies (e.g., oral or
 intravenous corticosteroids) due to their potential to obscure immunogenicity or
 immunoreactivity to Test RBCs.

 -  Subjects donating blood for any other purpose

 -  Subjects who have received blood transfusion within the previous year

 -  Subjects who are enrolled in another study.