Journal article

Tetramer Immunization and Selection Followed by CELLISA Screening to Generate Monoclonal Antibodies against the Mouse Cytomegalovirus m12 Immunoevasin

Oscar A Aguilar, Miho Tanaka, Gautham R Balaji, Richard Berry, Jamie Rossjohn, Lewis L Lanier, James R Carlyle

Journal of Immunology | AMER ASSOC IMMUNOLOGISTS | Published : 2020

Abstract

The generation of reliable mAb of unique and desired specificities serves as a valuable technology to study protein expression and function. However, standard approaches to mAb generation usually involve large-scale protein purification and intensive screening. In this study, we describe an optimized high-throughput proof-of-principle method for the expanded generation, enrichment, and screening of mouse hybridomas secreting mAb specific for a protein of interest. Briefly, we demonstrate that small amounts of a biotinylated protein of interest can be used to generate tetramers for use as prime-boost immunogens, followed by selective enrichment of Ag-specific B cells by magnetic sorting using..

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University of Melbourne Researchers

Grants

Awarded by Canadian Institutes of Health Research Operating Grants


Awarded by Burroughs Wellcome Fund (BWF) Investigator in the Pathogenesis of Infectious Disease Award


Awarded by Australian Research Council


Awarded by National Health and Medical Research Council of Australia


Funding Acknowledgements

This work was supported by Canadian Institutes of Health Research Operating Grants MOP 106491 and PJT 159450 (to J.R.C.) and by Burroughs Wellcome Fund (BWF) Investigator in the Pathogenesis of Infectious Disease Award 1007761 (to J.R.C.) and a BWF Postdoctoral Enrichment Program award (to O.A.A). O.A.A. is a Cancer Research Institute Irvington Fellow supported by the Cancer Research Institute. O.A.A. and L.L.L. are supported by the Parker Institute for Cancer Immunotherapy. R. is a recipient of Australian Research Council Laureate Fellowship FL160100049. R.B. is a recipient of National Health and Medical Research Council of Australia Career Development Award APP1109901.