Development and identification of monoclonal antibodies against c-erbB-2 p185 intracellular domain

To develop and identify the monoclonal antibodies (mAb) against c-erbB-2 p185 intracellular domain for detection of c-erbB-2 protein overexpression in breast tumor cells.

BALB/C mice were immunized with a synthesized p185 peptide of intracellular domain. The biological characteristics and immunoactivities were identified by different techniques.

Three hybridoma cell strains secreting mAbs to c-erbB-2 protein were established and one of the mAbs, No. 035-E61 was tested for c-erbB-2 protein immunostaining on 39 breast carcinoma sections, 30 breast fibroadenoma sections and 16 sections from various normal organs. The results showed that the positive detection frequency of protein expression was 26% (10/39) in breast cancer, none (0/30) in fibroadenoma and 3 sections from normal organs were positively stained. The consistency between the 035-E61 and the DAKO reagent approved by FDA was 74%.

No. 035-E61 mAb can specifically recognize the p185 intracellular domain and may be useful in guiding clinical Herceptin treatment.