Anti-mPD-1-mIgG1e3 InvivoFit™ is a recombinant monoclonal antibody (mAb) featuring the variable region of the previously described anti-mPD-1 RMP1-14 mAb [1, 2]. The original RMP1-14 hybridoma was obtained by immunizing rats with cells expressing murine programmed cell death 1 (mPD-1; also known as CD279). The use of xenogeneic sequences (i.e. rat origin) for mAbs renders them immunogenic upon injection in mice [3]. Moreover, special attention should apply to mAbs targeting the PD-1/PD-L1 axis, as repeated injections of xenogeneic anti-PD-1 or anti-PD-L1 in tumor-bearing mice were shown to induce fatal hypersensitivity reactions [4]. To overcome this issue, Anti-mPD-1-mIgG1e3 InvivoFit™ was generated by recombinant DNA technology so that it is ~65% murine. It contains the constant region of mouse IgG1 with a D265A point mutation (replacement of aspartic acid by alanine at position 265) resulting in the complete loss of cytolytic effector function [2, 5]. Anti-mPD-1-mIgG1e3 is provided in an InvivoFit™ grade, a high-quality standard specifically adapted to in vivo studies.
Key features of Anti-mPD-1-mIgG1e3 InvivoFit™:
- Derives from the RMP1-14 clone, rat IgG2a,κ
- Features mouse IgG1e3 isotype (constant region)
- mIgG1e3 (IgG1 with a D265A point mutation) is effectorless
- Blocks the murine PD-1 receptor without causing T cell depletion
- Guaranteed sterile, endotoxin level < 1 EU/mg
- Specifically designed for in vivo studies in mice
- Low aggregation < 5%
- Produced in animal-free facilities and defined media
Anti-mPD -1-mIgG1e3 InvivoFit™ is produced in Chinese hamster ovary (CHO) cells, purified by affinity chromatography with protein A, and its binding is validated by flow cytometry and ELISA.
References:
- Ribas A. & Wolchock J.D., 2018. Cancer immunotherapy using checkpoint blockade. Science. 359:1350-55.
- Yamazaki T. et al., 2005. Blockade of B7-H1 on macrophages suppresses CD4+ T cell proliferation by augmenting IFN-gamma-induced nitric oxide production. J Immunol. 175(3):1586-92.
- Brüggemann M. et al., 1989. The immunogenicity of chimeric antibodies. J. Exp. Med. 170:2153-2157.
- Mall C. et al., 2016. Repeated PD-1/PD-L1 monoclonal antibody administration induces fatal xenogeneic hypersensitivity reactions in a murine model of breast cancer. Oncoimmunology. 5(2):e1075114.
- Baudino L. et al., 2008. Crucial role of aspartic acid at position 265 in the CH2 domain for murine IgG2a and IgG2b Fc-associated effector functions. J. Immunol. 181(9):6664-9.