Universität zu Köln

Gieselmann, Lutz, Kreer, Christoph ORCID: 0000-0002-9140-9850, Ercanoglu, Meryem Seda, Lehnen, Nathalie, Zehner, Matthias, Schommers, Philipp ORCID: 0000-0003-3375-6800, Potthoff, Julian, Gruell, Henning and Klein, Florian (2021). Effective high-throughput isolation of fully human antibodies targeting infectious pathogens. Nat. Protoc., 16 (7). S. 3639 - 3672. BERLIN: NATURE PORTFOLIO. ISSN 1750-2799

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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. The authors provide a highly efficient high-throughput protocol for the isolation of potent pathogen-specific antibodies.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Gieselmann, Lutz UNSPECIFIED UNSPECIFIED UNSPECIFIED Kreer, Christoph UNSPECIFIED orcid.org/0000-0002-9140-9850 UNSPECIFIED Ercanoglu, Meryem Seda UNSPECIFIED UNSPECIFIED UNSPECIFIED Lehnen, Nathalie UNSPECIFIED UNSPECIFIED UNSPECIFIED Zehner, Matthias UNSPECIFIED UNSPECIFIED UNSPECIFIED Schommers, Philipp UNSPECIFIED orcid.org/0000-0003-3375-6800 UNSPECIFIED Potthoff, Julian UNSPECIFIED UNSPECIFIED UNSPECIFIED Gruell, Henning UNSPECIFIED UNSPECIFIED UNSPECIFIED Klein, Florian UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-571630
DOI:	10.1038/s41596-021-00554-w
Journal or Publication Title:	Nat. Protoc.
Volume:	16
Number:	7
Page Range:	S. 3639 - 3672
Date:	2021
Publisher:	NATURE PORTFOLIO
Place of Publication:	BERLIN
ISSN:	1750-2799
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language BROADLY NEUTRALIZING ANTIBODIES; HUMAN MONOCLONAL-ANTIBODIES; B-CELLS; VIRUS; POTENT; HIV; GENERATION; PROTECTION; IMMUNOGLOBULIN; SINGLE Multiple languages Biochemical Research Methods Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/57163