Recombinant mouse mAb against gp75 tumor-associated antigen for in vivo use

InvivoGen’s Anti-mgp75-mIgG2a is free of aberrant light chains

Anti-mgp75-mIgG2a InvivoFit™ is a recombinant mouse IgG2a monoclonal antibody (mAb) based on the previously described TA99 mAb that reacts against both human and mouse gp75 antigens [1]. The TA99 mAb has a sequence that is 100% murine (constant and variable regions), thus ensuring minimal immunogenicity upon repeated injections into mice. This mAb efficiently induces protective immunity against B16F10 melanoma, either subcutaneous or metastatic [2]. Of note, the mIgG2a isotype was shown to be the most potent at inducing antibody-dependent cellular cytotoxicity compared to mIgG1, mIgG2b, and mIgG3 [3]. As opposed to the original hybridoma-based production which contains a mixed population of anti-gp75 antibodies featuring correct or aberrant light chains [4], InvivoGen produces the recombinant anti-mgp75-mIgG2a mAb using Chinese hamster ovary (CHO) cells.Anti-mgp75-mIgG2a is purely monoclonal and is provided in an InvivoFit™ grade, a high-quality standard specifically adapted toin vivo studies.

Anti-mgp75-mIgG2a InvivoFit™ is specifically adapted for in vivo use in mice:

• Sequence is 100% murine
• Free from non-relevant mAbs found in hybridoma-based productions
• Guaranteed sterile, endotoxin level < 1 EU/mg
• Suitable for parenteral delivery in mice (azide-free)
• Low aggregation < 5%
• Produced in both animal-free facilities and defined media

Anti-mgp75-mIgG2a InvivoFit™ is purified by affinity chromatography with protein A. Its binding to mouse gp-75 is validated using ELISA.

References:

1. Thomson T.M. et al., 1985. Pigmentation-associated glycoprotein of human melanomas and melanocytes: definition with a mouse monoclonal antibody. J Invest Dermatol. 85(2):169-74.2. Hara I. et al., 1995. Implicating a role for immune recognition of self in tumor rejection: passive immunization against the brown locus protein. J Exp Med. 182(5):1609-14.3. Nimmerjahn F. & Ravetch J.V., 2005. Divergent immunoglobulin g subclass activity through selective Fc receptor binding. Science. 310(5753):1510-2.4. Boross P. et al., 2014. Anti-tumor activity of human IgG1 anti-gp75 TA99 mAb against B16F10 melanoma in human FcgammaRI transgenic mice. Immunol Lett. 160(2):151-7.

Figures

ELISA binding of Anti-mgp75-mIgG2a InvivoFit™ to recombinant murine  gp75 protein.

A dilution series of Anti-mgp75-mIgG2a InvivoFit™ and anti-mgp75 (clone TA99) was used for the detection of coated mgp75 (mTYRP1) antigen at 2 µg/ml. Detection of bound antibody was performed using an HRP-conjugated anti-murine IgG2a chain secondary antibody. HRP activity was assessed by a colorimetric assay using Tetramethylbenzidine (TMB) substrate. Optical density was measured at 490 nm. Data are shown as percentage absorbance.

Specifications

Specificity: Targets cells expressing murine gp75

Formulation: Lyophilized from 0.2 μm filtered solution in 150 mM sodium chloride, 20 mM sodium phosphate buffer with 5% saccharose​

Clonality: Monoclonal antibody

Isotype: Murine IgG2a, kappa 

Source: CHO cells

Purity: Purified by affinity chromatography with protein A

Tested applications: ELISA

Quality control:

• Binding confirmed using ELISA
• The complete sequence of this antibody has been verified
• < 5% aggregates (confirmed by size exclusion chromatography)
• Guaranteed sterile and <1 EU/mg (determined by the LAL assay)

Contents

Anti-mgp75-mIgG2a InvivoFit™ is provided sterile, endotoxin-free, azide-free and lyophilized.  

This product is available in two pack sizes:

• mgp75-mab10-1 : 1 mg
• mgp75-mab10-10 : 10 mg

 Product is shipped at room temperature.

 Store lyophilized antibody at -20 °C.

 Lyophilized product is stable for at least 1 year

 Avoid repeated freeze-thaw cycles.

Details

Glycoprotein 75 (gp75) is a tumor-associated antigen specifically located within the membrane of melanosomes; the organelles producing melanin in melanocytes. This 75-kDa polypeptide is the mature form of tyrosinase-related protein  1 (TYRP1; also known as MEL-5).This enzyme is the most abundant glycoprotein synthesized by pigmented melanocytes and melanomas. It is involved in melanin synthesis as well as the proliferation and cell death of melanocytes. Importantly, gp75 is a target of choice for antibody-based immunotherapy against primary and metastatic melanoma.