Anti-HE4 Antibody: Cancer Diagnosis Breakthrough | AtaGenix

A collaborative study developed a high-yield mammalian expression system for HE4 and a high-affinity mAb (9C3), with AtaGenix providing recombinant protein expression, hybridoma preparation, and in vivo validation.

Background

Human epididymis protein-4 (HE4) is a glycoprotein biomarker for ovarian, lung, breast, and pancreatic cancers. Traditional markers like CA125 lack specificity, making high-affinity anti-HE4 antibodies essential for early diagnosis and therapy.

Methods and AtaGenix’s Role

A high-yield system produced recombinant HE4 in HEK293 culture (400 mg/L). AtaGenix supplied HE4 cDNA, plasmid vectors, E. coli DH5α, HEK293 cells, and SPF animal facilities for in vivo experiments.

Purified HE4 was used for hybridoma generation. AtaGenix’s hybridoma platform screened clones, selecting 9C3 for high titer. AtaGenix also provided HRP-conjugated secondary antibodies for assays.

Key Findings

1) Functional Recombinant HE4 Protein

Purified HE4 promoted proliferation and cell cycle progression in SKOV3 cells, confirming its antigen suitability.

2) Generation of Anti-HE4 Antibody 9C3

Hybridoma screening yielded 9C3 with high titer. Ascites produced 24 mg purified antibody (>95% purity). Biacore: KD = 5.5×10⁻¹² M; DSC: Tm = 73°C.

3) Specificity and Applications

Western blot, IHC, and IF confirmed 9C3’s specificity for HE4 in proteins, cell lines, and tissues, making it ideal for diagnostics.

Significance

Combining mammalian expression, hybridoma tech, and characterization, the study created a high-affinity anti-HE4 antibody. AtaGenix’s capabilities in antigen production and antibody discovery provide a model for biomarker-targeted development.

9C3 could enhance early ovarian cancer detection and oncology diagnostics.

References

1. Shen Y, Wang Y, Jiang X, et al. Preparation and characterization of a high-affinity monoclonal antibody against human epididymis protein-4. Elsevier, 2017.