Mammalian display to secretion switchable libraries for antibody preselection and high throughput functional screening

Recently, there has been a co-evolution of mammalian libraries and diverse microfluidic approaches for therapeutic antibody hit discovery. Mammalian libraries enable the preservation of full immune repertoires, produce hit candidates in final format and facilitate broad combinatorial bispecific anti...

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Main Authors: Ramona Gaa, Hannah Melina Mayer, Daniela Noack, Kavita Kumari, Ralf Guenther, Shang-Pu Tsai, Qingyong Ji, Achim Doerner
Format: Article
Language:English
Published: Taylor & Francis Group 2023-12-01
Series:mAbs
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Online Access:https://www.tandfonline.com/doi/10.1080/19420862.2023.2251190
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author Ramona Gaa
Hannah Melina Mayer
Daniela Noack
Kavita Kumari
Ralf Guenther
Shang-Pu Tsai
Qingyong Ji
Achim Doerner
author_facet Ramona Gaa
Hannah Melina Mayer
Daniela Noack
Kavita Kumari
Ralf Guenther
Shang-Pu Tsai
Qingyong Ji
Achim Doerner
author_sort Ramona Gaa
collection DOAJ
description Recently, there has been a co-evolution of mammalian libraries and diverse microfluidic approaches for therapeutic antibody hit discovery. Mammalian libraries enable the preservation of full immune repertoires, produce hit candidates in final format and facilitate broad combinatorial bispecific antibody screening, while several available microfluidic methodologies offer opportunities for rapid high-content screens. Here, we report proof-of-concept studies exploring the potential of combining microfluidic technologies with mammalian libraries for antibody discovery. First, antibody secretion, target co-expression and integration of appropriate reporter cell lines enabled the selection of in-trans acting agonistic bispecific antibodies. Second, a functional screen for internalization was established and comparison of autocrine versus co-encapsulation setups highlighted the advantages of an autocrine one cell approach. Third, synchronization of antibody-secreting cells prior to microfluidic screens reduced assay variability. Furthermore, a display to secretion switchable system was developed and applied for pre-enrichment of antibody clones with high manufacturability in conjunction with subsequent screening for functional properties. These case studies demonstrate the system’s feasibility and may serve as basis for further development of integrated workflows combining manufacturability sorting and functional screens for the identification of optimal therapeutic antibody candidates.
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1942-0870
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spelling doaj-art-1d7d7b1dbf634c3f954356ece3b083e82025-08-20T02:50:17ZengTaylor & Francis GroupmAbs1942-08621942-08702023-12-0115110.1080/19420862.2023.2251190Mammalian display to secretion switchable libraries for antibody preselection and high throughput functional screeningRamona Gaa0Hannah Melina Mayer1Daniela Noack2Kavita Kumari3Ralf Guenther4Shang-Pu Tsai5Qingyong Ji6Achim Doerner7NBE Technologies, Merck KGaA, Darmstadt, GermanyNBE Technologies, Merck KGaA, Darmstadt, GermanyNBE Technologies, Merck KGaA, Darmstadt, GermanyDiscovery Biology, Syngene International, Bangalore, IndiaNBE Technologies, Merck KGaA, Darmstadt, GermanyNBE Technologies, EMD Serono, Billerica, MA, USANBE Technologies, EMD Serono, Billerica, MA, USANBE Technologies, Merck KGaA, Darmstadt, GermanyRecently, there has been a co-evolution of mammalian libraries and diverse microfluidic approaches for therapeutic antibody hit discovery. Mammalian libraries enable the preservation of full immune repertoires, produce hit candidates in final format and facilitate broad combinatorial bispecific antibody screening, while several available microfluidic methodologies offer opportunities for rapid high-content screens. Here, we report proof-of-concept studies exploring the potential of combining microfluidic technologies with mammalian libraries for antibody discovery. First, antibody secretion, target co-expression and integration of appropriate reporter cell lines enabled the selection of in-trans acting agonistic bispecific antibodies. Second, a functional screen for internalization was established and comparison of autocrine versus co-encapsulation setups highlighted the advantages of an autocrine one cell approach. Third, synchronization of antibody-secreting cells prior to microfluidic screens reduced assay variability. Furthermore, a display to secretion switchable system was developed and applied for pre-enrichment of antibody clones with high manufacturability in conjunction with subsequent screening for functional properties. These case studies demonstrate the system’s feasibility and may serve as basis for further development of integrated workflows combining manufacturability sorting and functional screens for the identification of optimal therapeutic antibody candidates.https://www.tandfonline.com/doi/10.1080/19420862.2023.2251190Antibodiesdisplay secretion switchfunction first screenmammalian librariesmicrofluidics
spellingShingle Ramona Gaa
Hannah Melina Mayer
Daniela Noack
Kavita Kumari
Ralf Guenther
Shang-Pu Tsai
Qingyong Ji
Achim Doerner
Mammalian display to secretion switchable libraries for antibody preselection and high throughput functional screening
mAbs
Antibodies
display secretion switch
function first screen
mammalian libraries
microfluidics
title Mammalian display to secretion switchable libraries for antibody preselection and high throughput functional screening
title_full Mammalian display to secretion switchable libraries for antibody preselection and high throughput functional screening
title_fullStr Mammalian display to secretion switchable libraries for antibody preselection and high throughput functional screening
title_full_unstemmed Mammalian display to secretion switchable libraries for antibody preselection and high throughput functional screening
title_short Mammalian display to secretion switchable libraries for antibody preselection and high throughput functional screening
title_sort mammalian display to secretion switchable libraries for antibody preselection and high throughput functional screening
topic Antibodies
display secretion switch
function first screen
mammalian libraries
microfluidics
url https://www.tandfonline.com/doi/10.1080/19420862.2023.2251190
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