Bogen, Jan P. ; Storka, Juliana ; Yanakieva, Desislava ; Fiebig, David ; Grzeschik, Julius ; Hock, Björn ; Kolmar, Harald (2024)
Isolation of Common Light Chain Antibodies from Immunized Chickens Using Yeast Biopanning and Fluorescence‐Activated Cell Sorting.
In: Biotechnology Journal : Systems and Synthetic Biology, Nanobiotech, Medicine, 2020, 16 (3)
doi: 10.26083/tuprints-00017449
Artikel, Zweitveröffentlichung, Verlagsversion

Kurzbeschreibung (Abstract)

The phylogenetic distance between chickens and humans accounts for a strong immune response and a broader epitope coverage compared to rodent immunization approaches. Here the authors report the isolation of common light chain (cLC)‐based chicken monoclonal antibodies from an anti‐epidermal growth factor receptor (EGFR) immune library utilizing yeast surface display in combination with yeast biopanning and fluorescence‐activated cell sorting (FACS). For the selection of high‐affinity antibodies, a yeast cell library presenting cLC‐comprising fragment antigen binding (Fab) fragments is panned against hEGFR‐overexpressing A431 cells. The resulting cell–cell‐complexes are sorted by FACS resulting in gradual enrichment of EGFR‐binding Fabs in three sorting rounds. The isolated antibodies share the same light chain and show high specificity for EGFR, resulting in selective binding to A431 cells with notable EC50 values. All identified antibodies show very good aggregation propensity profiles and thermostabilities. Additionally, epitope binning demonstrates that these cLC antibodies cover a broad epitope space. Isolation of antibodies from immunized chickens by yeast cell biopanning makes an addition to the repertoire of methods for antibody library screening, paving the way for the generation of cLC‐based bispecific antibodies against native mammalian receptors.

Typ des Eintrags:	Artikel
Erschienen:	2024
Autor(en):	Bogen, Jan P. ; Storka, Juliana ; Yanakieva, Desislava ; Fiebig, David ; Grzeschik, Julius ; Hock, Björn ; Kolmar, Harald
Art des Eintrags:	Zweitveröffentlichung
Titel:	Isolation of Common Light Chain Antibodies from Immunized Chickens Using Yeast Biopanning and Fluorescence‐Activated Cell Sorting
Sprache:	Englisch
Publikationsjahr:	30 Januar 2024
Ort:	Darmstadt
Publikationsdatum der Erstveröffentlichung:	2020
Ort der Erstveröffentlichung:	Weinheim
Verlag:	Wiley-VCH
Titel der Zeitschrift, Zeitung oder Schriftenreihe:	Biotechnology Journal : Systems and Synthetic Biology, Nanobiotech, Medicine
Jahrgang/Volume einer Zeitschrift:	16
(Heft-)Nummer:	3
Kollation:	10 Seiten
DOI:	10.26083/tuprints-00017449
URL / URN:	https://tuprints.ulb.tu-darmstadt.de/17449
Zugehörige Links:	Verlags-DOI
Herkunft:	Zweitveröffentlichung DeepGreen
Kurzbeschreibung (Abstract):	The phylogenetic distance between chickens and humans accounts for a strong immune response and a broader epitope coverage compared to rodent immunization approaches. Here the authors report the isolation of common light chain (cLC)‐based chicken monoclonal antibodies from an anti‐epidermal growth factor receptor (EGFR) immune library utilizing yeast surface display in combination with yeast biopanning and fluorescence‐activated cell sorting (FACS). For the selection of high‐affinity antibodies, a yeast cell library presenting cLC‐comprising fragment antigen binding (Fab) fragments is panned against hEGFR‐overexpressing A431 cells. The resulting cell–cell‐complexes are sorted by FACS resulting in gradual enrichment of EGFR‐binding Fabs in three sorting rounds. The isolated antibodies share the same light chain and show high specificity for EGFR, resulting in selective binding to A431 cells with notable EC50 values. All identified antibodies show very good aggregation propensity profiles and thermostabilities. Additionally, epitope binning demonstrates that these cLC antibodies cover a broad epitope space. Isolation of antibodies from immunized chickens by yeast cell biopanning makes an addition to the repertoire of methods for antibody library screening, paving the way for the generation of cLC‐based bispecific antibodies against native mammalian receptors.
Alternatives oder übersetztes Abstract:	Alternatives Abstract Sprache A common light chain yeast surface display library derived from immunized chickens is generated and high affine antibody variants are isolated using yeast biopanning in combination with fluorescence-activated cell sorting. Resulting antibodies address a broad epitope space and exhibit favorable aggregation propensities and thermal stabilities while sharing a single light chain. Englisch
Freie Schlagworte:	antibody discovery, cell panning, common light chain, fluorescence‐activated cell sorting, yeast display
ID-Nummer:	Artikel-ID: 2000240
Status:	Verlagsversion
URN:	urn:nbn:de:tuda-tuprints-174498
Sachgruppe der Dewey Dezimalklassifikatin (DDC):	500 Naturwissenschaften und Mathematik > 540 Chemie 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie
Fachbereich(e)/-gebiet(e):	07 Fachbereich Chemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut > Fachgebiet Biochemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut
Hinterlegungsdatum:	30 Jan 2024 13:26
Letzte Änderung:	31 Jan 2024 06:58
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Verfügbare Versionen dieses Eintrags

• Isolation of Common Light Chain Antibodies from Immunized Chickens Using Yeast Biopanning and Fluorescence‐Activated Cell Sorting. (deposited 30 Jan 2024 13:26) [Gegenwärtig angezeigt]
  • Isolation of common light chain antibodies from immunized chickens using yeast biopanning and fluorescence‐activated cell sorting. (deposited 31 Jan 2024 06:59)

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