Automated pipeline for rapid production and screening of HIV-specific monoclonal antibodies using pichia pastoris.

Autor: Shah KA; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts., Clark JJ; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts., Goods BA; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts., Politano TJ; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts., Mozdzierz NJ; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts., Zimnisky RM; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts., Leeson RL; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts., Love JC; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts. clove@mit.edu.; MIT Center for Biomedical Innovation, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts. clove@mit.edu., Love KR; Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts. kerryluv@mit.edu.; MIT Center for Biomedical Innovation, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts. kerryluv@mit.edu.
Jazyk: angličtina
Zdroj: Biotechnology and bioengineering [Biotechnol Bioeng] 2015 Dec; Vol. 112 (12), pp. 2624-9. Date of Electronic Publication: 2015 Jul 31.
DOI: 10.1002/bit.25663
Abstrakt: Monoclonal antibodies (mAbs) that bind and neutralize human pathogens have great therapeutic potential. Advances in automated screening and liquid handling have resulted in the ability to discover antigen-specific antibodies either directly from human blood or from various combinatorial libraries (phage, bacteria, or yeast). There remain, however, bottlenecks in the cloning, expression and evaluation of such lead antibodies identified in primary screens that hinder high-throughput screening. As such, "hit-to-lead identification" remains both expensive and time-consuming. By combining the advantages of overlap extension PCR (OE-PCR) and a genetically stable yet easily manipulatable microbial expression host Pichia pastoris, we have developed an automated pipeline for the rapid production and screening of full-length antigen-specific mAbs. Here, we demonstrate the speed, feasibility and cost-effectiveness of our approach by generating several broadly neutralizing antibodies against human immunodeficiency virus (HIV).
(© 2015 Wiley Periodicals, Inc.)
Databáze: MEDLINE
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