Biotin for red blood cells labeling

Paresh_jain@BD.com Paresh_jain at BD.com
Tue Nov 8 21:21:48 EST 2005


Hello Grigoriy

You might find the following information useful. This information was 
submitted by a scientist from JNU, Delhi.

regards

Dr. Paresh Jain
Scientific Advisor
BD Biosciences, 
India.

Dear sir
how r u ? 
three daily doses of Biotin X NHS ( calbiochem, Cat no. 203188) injecte 
Intravenously gives 100 % biotin labeling of blood erythrocytes in mice. 
for detail i giving one reference.
Biotin labeling as an alternative nonradioactive approach to determination 
of red cell survival.
Annal Hematol 1993, vol 67, 81-87.
In this paper detail method of erythrocyte biotin labeling has been given
 with regards
sanjay




"Guy Hermans" <Guy.Hermans at ablynx.com>
11/08/2005 01:07 PM
 
        To:     Cytometry Mailing List <cytometry at flowcyt.cyto.purdue.edu>
        cc:     (bcc: Paresh jain/IND/BDX)
        Subject:        RE: Biotin for red blood cells labeling


Hi Grigoriy,
 
I fail to see why getting total cell population labeling is required to 
determine that populaqtion's half-life. Better get high intensity staining 
of a subset with a reagent which won't interfere with turnover than 
getting total labeling with something which might change the observed 
parameter - and I'm not so sure biotin is the best choice in that respect 
(I may be wrong here).
 
I'd recommend labeling with CFSE if that's possible, or ex-vivo label 
packed cells with CFSE and reintroduce them via tail vein. This label is 
"invisible" biochemically on the outside of the cells and is therefore 
less likely to change halflife. Also, it allows direct measurement of 
fluorescence, so any additional time spent during labeling in this 
alternative protocol would be regained during sampling/analysis.
 
In any case: I'd suggest introducing the label (biotin or otherwise) 
intravenously (tail vein) to get better/faster distribution than 
retroorbital.
 
Good luck,
 
Guy
 
 




Next generation therapeutic antibodies



Guy Hermans, PhD
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-----Original Message-----
From: Losyev, Grigoriy [mailto:Grigoriy.Losyev at childrens.harvard.edu] 
Sent: Friday, November 04, 2005 6:38 PM
To: Cytometry Mailing List
Subject: Biotin for red blood cells labeling

Dear Flowers
 
Our current project is labeling RBCs with 1.5 mg NHS PEO4 biotin injected 
retro-orbitally in 3 doses over 24 hours and then watching RBC aging 
through time. 
After labeling RBCs in mouse’s blood stream we have drawn blood twice a 
week and stain RBCs with Alexa 488 SA.  Then we pass cells trough 
flowcytometer. We are measuring percent of stained cells. During the time 
course it goes down.
 
My question is related to biotin quality. We never were able to label 90% 
of RBC at the initial point. Does it depend on biotin? Is there a better 
grade for biotin?
Could somebody suggest what could improve initial labeling of RBCs?
 
Thanks 
 
 
Grigoriy Losyev
Division of Hematology/Oncology
Flow Cytometry
Children’s Hospital Boston
Karp Family Research Facilities, 08010
300 Longwood Ave 
Boston MA 02115 
Phone- 617.919.2127
Fax- 617.730.0934 
 

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