Generation of a cost-effective cell line for support of high-throughput isolation of primary human B cells and monoclonal neutralizing antibodies

• Authors: Rachael E. Whaley, Fred Hutchinson Cancer Research Center; Sarah Ameny, Fred Hutchinson Cancer Research Center; Tanvi Arkatkar, Fred Hutchinson Cancer Research Center; Aaron Seese, Fred Hutchinson Cancer Research Center; Abigail Wall, Fred Hutchinson Cancer Research Center; Iram Khan, Seattle Children's Hospital; Joseph J. Carter, Fred Hutchinson Cancer Research Center; Erin Scherer, Emory University; David J. Rawlings, Seattle Children's Hospital; Denise A. Galloway, Fred Hutchinson Cancer Research Center; M. Juliana McElrath, Fred Hutchinson Cancer Research Center; Kristen W. Cohen, Fred Hutchinson Cancer Research Center; Andrew T. McGuire, Fred Hutchinson Cancer Research Center
• Journal Title: JOURNAL OF IMMUNOLOGICAL METHODS
• Volume: Volume 488
• Publisher: ELSEVIER | 2021-01-01, Pages 112901-112901
• Type of Work: Article | Final Publisher PDF
• Publisher DOI: http://doi.org/10.1016/j.jim.2020.112901
• Subjects: Health Sciences, Immunology; Chemistry, Biochemistry; Biology, Molecular; Biology, Virology; Health Sciences, Oncology
• Keywords: Science & Technology; Life Sciences & Biomedicine; Biochemical Research Methods; Immunology; Biochemistry & Molecular Biology; B cells; Antibodies; High-throughput; Screening; Human papillomavirus; Cytokines; RESPIRATORY SYNDROME CORONAVIRUS; POTENT NEUTRALIZATION; IMMUNODEFICIENCY-VIRUS; PASSIVE TRANSFER; RECEPTOR; MEMORY; HIV-1; PROTECTION; INFECTION; BINDING

Abstract

The isolation of human monoclonal antibodies (mAbs) arising from natural infection with human pathogens has proven to be a powerful technology, facilitating the understanding of the host response to infection at a molecular level. mAbs can reveal sites of vulnerability on pathogens and illuminate the biological function of the antigenic targets. Moreover, mAbs have the potential to be used directly for therapeutic applications such as passive delivery to prevent infection in susceptible target populations, and as treatment of established infection. The isolation of antigen-specific B cells from vaccine trials can also assist in deciphering whether the desired B cells are being targeted by a given vaccine. Several different processes have been developed to isolate mAbs, but all are generally labor-intensive and result in varying degrees of efficiency. Here, we describe the development of a cost-effective feeder cell line that stably expresses CD40-ligand, interleukin-2 and interleukin-21. Sorting of single B cells onto a layer of irradiated feeder cells sustained antibody production that permits functional screening of secreted antibodies in a manner that enables subsequent recovery of B cells for recombinant antibody cloning. As a proof of concept, we show that this approach can be used to isolate B cells that secrete antibodies that neutralize human papilloma virus (HPV) from participants of an HPV vaccine study.