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Coupling brain perfusion screens and next generation sequencing to identify blood–brain barrier binding antibodies
Author(s) -
Stutz Charles C.,
Georgieva Julia V.,
Shusta Eric V.
Publication year - 2018
Publication title -
aiche journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.958
H-Index - 167
eISSN - 1547-5905
pISSN - 0001-1541
DOI - 10.1002/aic.16360
Subject(s) - coupling (piping) , blood–brain barrier , antibody , perfusion , biophysics , neuroscience , computational biology , chemistry , medicine , biology , materials science , immunology , cardiology , central nervous system , metallurgy
Antibodies that target the blood–brain barrier (BBB) in vivo are of particular interest for the treatment of neurological diseases. Here, we screened a phage display single‐chain antibody (scFv) library by brain perfusion in an attempt to isolate scFv that target the rat BBB. After four rounds of screening, the resulting antibody pool remained highly complex and discrete clonal sampling did not identify any scFvs capable of binding to the rat BBB. Thus, the heavy chain CDR3 in the resulting pools was subjected to next generation sequencing (NGS), and the resulting data was used to identify 12 scFv clones that were of high abundance and/or enriched from Round 3 to 4, signifying potential hits. Of these, two scFv, denoted scFv 4 and scFv 40, were identified that bound the rat BBB. Neither of these scFvs was identified by discrete sampling, motivating NGS as a tool to identify lead antibodies from complex in vivo screens. © 2018 American Institute of Chemical Engineers AIChE J , 64: 4229–4236, 2018