Generation and characterization of genetically stable heterohybridomas producing foot-and-mouth disease virus-specific porcine monoclonal antibodies

Michael C Puckette; Erica Martel; Jacob Rutherford; José Barrera; William Hurtle; Melia Pisano; Lauren Martignette; Mariceny Zurita; John G Neilan; Chungwon J Chung

doi:10.1016/j.jim.2020.112873

Generation and characterization of genetically stable heterohybridomas producing foot-and-mouth disease virus-specific porcine monoclonal antibodies

J Immunol Methods. 2020 Dec:487:112873. doi: 10.1016/j.jim.2020.112873. Epub 2020 Sep 28.

Authors

Affiliations

¹ U.S. Department of Homeland Security Science & Technology Directorate, Plum Island Animal Disease Center, Greenport, NY, USA. Electronic address: Michael.puckette@st.dhs.gov.
² Oak Ridge Institute for Science and Education, Plum Island Animal Disease Center Research Participation Program, Oak Ridge, TN, USA. Electronic address: Martel52@rowan.edu.
³ Oak Ridge Institute for Science and Education, Plum Island Animal Disease Center Research Participation Program, Oak Ridge, TN, USA. Electronic address: jtruther@exchange.tamu.edu.
⁴ Leidos, Plum Island Animal Disease Center, Greenport, NY, USA. Electronic address: Jose.Barrera@st.dhs.gov.
⁵ U.S. Department of Homeland Security Science & Technology Directorate, Plum Island Animal Disease Center, Greenport, NY, USA. Electronic address: William.Hurtle@st.dhs.gov.
⁶ Leidos, Plum Island Animal Disease Center, Greenport, NY, USA.
⁷ Leidos, Plum Island Animal Disease Center, Greenport, NY, USA. Electronic address: lauren.martignette@st.dhs.gov.
⁸ Leidos, Plum Island Animal Disease Center, Greenport, NY, USA. Electronic address: mariceny.zurita@st.dhs.gov.
⁹ U.S. Department of Homeland Security Science & Technology Directorate, Plum Island Animal Disease Center, Greenport, NY, USA. Electronic address: john.neilan@st.dhs.gov.
¹⁰ U.S. Department of Homeland Security Science & Technology Directorate, Plum Island Animal Disease Center, Greenport, NY, USA. Electronic address: Chungwon.chung@usda.gov.

PMID: 32998052
DOI: 10.1016/j.jim.2020.112873

Abstract

This report covers the methodology for generation of stable heterohybridoma clones producing Foot-and-mouth disease virus (FMDV) reactive porcine monoclonal antibodies (mAbs). Swine received five inoculations of an inactivated O1 Manisa FMDV vaccine prior to the harvest of splenocytes. Due to the lack of a species-specific hybridoma fusion partner, the Sp2/0 murine myeloma cell line was utilized for the formation of porcine-murine heterohybridoma clones. Twenty-nine FMDV-reactive parental clones were generated. Following sub-cloning and monitoring of reactivity over 20 serial passages, eleven subclones derived from unique parental origins were characterized and are reported herein. This methodology demonstrated the production of porcine mAbs by fusion of porcine splenocytes from immunized pigs with murine myeloma cells to generate heterohybridomas. The porcine immune response may differ from the murine immune response in relation to recognized epitopes. Therefore, application of this methodology may provide valuable resources for swine immunology and enhance the understanding of the mechanisms for antibody based protection from diseases in swine.

Keywords: Foot-and-mouth disease virus; Heterohybridoma; Hybridoma; Monoclonal antibody; Porcine immunoglobulins.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Antibodies, Monoclonal / biosynthesis*
Antibodies, Monoclonal / genetics
Antibodies, Monoclonal / immunology
Antibodies, Neutralizing / biosynthesis*
Antibodies, Neutralizing / genetics
Antibodies, Neutralizing / immunology
Antibody Specificity
B-Lymphocytes / immunology
Cell Line
Cloning, Molecular
Foot-and-Mouth Disease / immunology
Foot-and-Mouth Disease / prevention & control*
Foot-and-Mouth Disease / virology
Foot-and-Mouth Disease Virus / immunology*
Hybridomas
Immunization
Mice
Spleen / immunology
Sus scrofa
Viral Vaccines / immunology
Viral Vaccines / pharmacology*

Substances

Antibodies, Monoclonal
Antibodies, Neutralizing
Viral Vaccines