Journal
AICHE JOURNAL
Volume 64, Issue 12, Pages 4229-4236Publisher
WILEY
DOI: 10.1002/aic.16360
Keywords
antibody engineering; antibody display
Categories
Funding
- National Institutes of Health [NS071513, NS099158]
- Defense Threat Reduction Agency [HDTRA1-15-1-0012]
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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. (c) 2018 American Institute of Chemical Engineers
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