Establishment of anti-asialo-GM1 rabbit monoclonal antibodies capable of reducing natural killer cell activity in mice

Tatsuji Kimura; Satoshi Ohta; Hiroshi Murayama

doi:10.1371/journal.pone.0292514

Establishment of anti-asialo-GM1 rabbit monoclonal antibodies capable of reducing natural killer cell activity in mice

PLoS One. 2023 Oct 9;18(10):e0292514. doi: 10.1371/journal.pone.0292514. eCollection 2023.

Authors

Tatsuji Kimura¹, Satoshi Ohta¹, Hiroshi Murayama¹

Affiliation

¹ Diagnostic Division, Yamasa Corporation, Choshi, Chiba, Japan.

Abstract

Rabbit anti-asialo-GM1 (ASGM1) serum or polyclonal antibodies can eliminate mouse splenic natural killer (NK) cell activity in vitro and in vivo. We developed rabbit monoclonal antibodies (mAbs) against ASGM1 using a single-cell analysis and isolation system. Five mAbs (GA109, GA115, GA116, GA131, and GA134) that were reactive to ASGM1 were isolated from the spleen lymphocytes of rabbits immunized with ASGM1. Enzyme-linked immunosorbent assay and thin-layer chromatography immunostaining results showed that the mAbs strongly reacted with ASGM1. Two mAbs (GA116 and GA134) reacted exclusively with ASGM1, whereas three mAbs (GA109, GA115, and GA131) showed slight or considerable cross-reactivity with GM1. The administration of the mAbs (4-20 μg) to BALB/c mice completely abolished NK cell activity in vivo. The anti-ASGM1 rabbit mAbs obtained in this study may provide a useful and reproducible tool for various future studies, such as depleting NK cell activity to enhance xenograft engraftment in mouse models.

Copyright: © 2023 Kimura et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Publication types

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

MeSH terms

Animals
Antibodies, Monoclonal
Enzyme-Linked Immunosorbent Assay
G(M1) Ganglioside*
Glycosphingolipids
Humans
Killer Cells, Natural*
Mice
Mice, Inbred BALB C
Mice, Inbred C57BL

Substances

G(M1) Ganglioside
Antibodies, Monoclonal
Glycosphingolipids

Grants and funding

The authors received no specific funding for this work. All work presented here was funded by Yamasa Corporation. The funder provided support in the form of salaries for authors (TK, SO, HM), but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the Author Contributions section.