Red Blood Cell Survival Following Transfusion in Infants
Phase: PHASE2
Conditions: Neonatal Anemia
Sponsor: John A Widness

Official Title: Red Blood Cell Survival Following Transfusion in Infants   
NCT00731588 
Date: 4/29/2017 

Change in Protocol: "RBC Survival in Adults with Prior Antibody Response to BioRBCs" (BB-IND 12990, #017) 
Page 1 of7 
Modifications of Prior Protocol Amendment: Change in Protocol "RBC Survival in 
Adults witlt Prior Antibody Response to BioRBCs" (BB-IND 12990 Serial Number 0008, 
submitted August 27, 2012) 
Note: Yellow highlighting indicates #0017 changes, i.e., since #0016 submission) 
Summary Addressing Specific FDA CRF 21 Sec 312.30 Protocol Amendments: (b) Changes in a 
protocol. 
This is a second revised protocol with minor changes replacing our prior Amended Protocol, 
"RBC Survival in Adults with Prior Antibody Res onse to BioRBCs" (BB-IND 12990 Serial 
Number 0008, submitted August 27, 2012 and its revision submitted July 19, 2016 as Serial 
iNumber 0016). Our goal as indicated below for this Phase 1 Study is to develop new 
knowledge that will improve the safety of administering BioRBCs to subjects who have 
developed anti-BioRBC antibodies following a prior BioRBC transfusion. We anticipate this is 
possible by administering BioRBCs in progressively increasing doses beginning with the lowest 
detectable 2.0 µg/mL BioRBC density ("BioRBC-2")-that we have been validating this past 
year-followed with subsequent progressive increases in BioRBC doses administered every 10-
28 d. Since the July 19, 2016 protocol modification, we have completed study of our first adulti 
study subject who received a single BioRBC dose of 2 µg/mL (results included below). In the 
cmTent #001 7 submission we propose a single minor modification of Protocol #0016 by 
combining the initial two lowest BioRBC doses in a single co-administration of both together to 
previously BioRBC sensitized adults, i.e., rather than as two se arate low BioRBC doses 
separated by 10 to 28 d. As indicated in the Rationale section below, we anticipate that his 
inor modification emains a potentially safer protocol than administering all five BioRBC 
doses (i.e., BioRBC-2, -6, -18, -54, and -162) at a single time because: 
(i)
A lower total BioRBC dose is likely possible because the design of the protocol has
been changed such that instead of the previous plan in our 2012 protocol amendment to
give all five autologous BioRBC density doses in a single BioRBC transfusion (i.e., 2,
6, 18, 54, and 128 µg/rnL all at once), the amended protocol calls for giving each of the
five BioRBC density doses one at a time in rogressively increasing density doses-
ow with the exception of the two lowest BioRBC density doses (2.0 and 6.0 µg/rnL) 
which will be co-administered at the same time. 
(ii)
Although no new antibody testing procedure will be added to our previous 2012
protocol, we will continue to apply the more sensitive IgG agglutination detection assay
we have developed to identify if and when BioRBC transfusion results in induction of
plasma anti-BioRBC antibodies. Compared to even a year ago, we are significantly
more experienced with and have developed improvement in this assay (1).
This modification is being concunently submitted to our IRB. Our initial FDA Amended 
Protocol was approved by our local IRB in 2012. 
A more complex Hypothesis (revised for #0017) that incorporates our latest observations. Upon 
re-exposure to autologous BioRBCs, adults who initially experienced an induced anti-BioRBC 
antibody response will demonstrate an anarnnestic immune response; the threshold for this 
response may be at densities as low as BioRBC-6, or even -2. However, the resultant anti­
BioRBC antibodies will not accelerate removal of the lowest BioRBC densities, i.e., BioRBC-2 
and -6. Thus, this hypothesis tacitly invokes two immunological thresholds: 1) one threshold for 
the initiation of an anarnnestic BioRBC antibody res onse; and 2) a second distinct and likely