Analysis of cerbB2 expression using a panel of 6 commercially available antibodies.
Results are presented of a study comparing cerbB2 (neu or Her2) expression as assessed immunohistochemically in breast neoplasia using a panel of 6 commercially available antibodies. The antibodies were examined utilizing conventional formalin fixed paraffin embedded tissue, and compared with molecular analysis of gene amplification. The aim was to determine the practical utility of each antibody, assessing ease of use, specific and non-specific staining characteristics, and expense, thus allowing a specific recommendation as to antibody of choice for immunohistochemical assessment of cerbB2 expression. Reassuringly, amongst the 38 breast lesions (36 carcinomas, 2 fibroadenomas) subjected to immunohistochemically (IHC) with the panel of 6 antibodies (Ab), no gross discrepancy of staining pattern was seen. Of the 38 cases, 10 were positive (26%), where at least one Ab demonstrated clear cytoplasmic membrane staining. Of a total of 45 breast lesions (43 carcinomas, 2 fibroadenomas), including all those examined by IHC, the total number of cases showing cerbB2 amplification by DNA analysis was 14 (31%). Using the DNA amplification as a base line for comparison, one Ab (No. 4) was found to stain 6 of the 14 cases of breast carcinoma that were assessed as showing amplification at the DNA level. Four Abs (1,3,5,6) stained 5 of these cases. However, Abs 3,4 and 6 displayed artefactual cytoplasmic staining (in the absence of membrane staining) that precluded the practical use of these reagents. Therefore, based on additional considerations of cost and ease of use, Ab No. 1 was finally chosen for recommendation from the 6 Ab panel.
Busmanis
I
I
Department of Anatomical Pathology, Royal Melbourne Hospital, Parkville, Victoria.
Feleppa
F
F
Jones
A
A
McGrath
K M
KM
Reed
R
R
Collins
J
J
Russell
I
I
Begley
C G
CG
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000906:Antibodies; D000911:Antibodies, Monoclonal; D015139:Blotting, Southern; D001943:Breast Neoplasms / Q000737:chemistry*; D002277:Carcinoma / Q000737:chemistry; D004273:DNA, Neoplasm / Q000032:analysis; D005260:Female; D018226:Fibroadenoma / Q000737:chemistry; D005784:Gene Amplification / Q000235:genetics; D006801:Humans; D007124:Immunoenzyme Techniques; D008875:Middle Aged; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis*
D000906:Antibodies; D000911:Antibodies, Monoclonal; D004273:DNA, Neoplasm; D018719:Erb-b2 Receptor Tyrosine Kinases