Effective high-throughput isolation of fully human antibodies targeting infectious pathogens.
Nat Protoc
; 16(7): 3639-3671, 2021 07.
Article
in English
| MEDLINE | ID: covidwho-1243308
ABSTRACT
As exemplified by the ongoing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, there is a strong demand for rapid high-throughput isolation pipelines to identify potent neutralizing antibodies for prevention and therapy of infectious diseases. However, despite substantial progress and extensive efforts, the identification and production of antigen-specific antibodies remains labor- and cost-intensive. We have advanced existing concepts to develop a highly efficient high-throughput protocol with proven application for the isolation of potent antigen-specific antibodies against human immunodeficiency virus 1, hepatitis C virus, human cytomegalovirus, Middle East respiratory syndrome coronavirus, SARS-CoV-2 and Ebola virus. It is based on computationally optimized multiplex primer sets (openPrimeR), which guarantee high coverage of even highly mutated immunoglobulin gene segments as well as on optimized antibody cloning and production strategies. Here, we provide the detailed protocol, which covers all critical steps from sample collection to antibody production within 12-14 d.
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Antibodies, Neutralizing
/
High-Throughput Screening Assays
/
COVID-19
Type of study:
Experimental Studies
Limits:
Humans
Language:
English
Journal:
Nat Protoc
Year:
2021
Document Type:
Article
Affiliation country:
S41596-021-00554-w
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