Glycobiology Advance Access originally published online on June 6, 2008
Glycobiology 2008 18(10):746; doi:10.1093/glycob/cwn049
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Published by Oxford University Press 2008.
Supplier-dependent antiglycan monoclonal antibody specificities: Comment on "High-throughput carbohydrate microarray profiling of 27 antibodies demonstrates widespread specificity problems"
2 Laboratory of Medicinal Chemistry, Center for Cancer Research, National Cancer Institute, 376 Boyles St. Bldg 376, Rm 109, Frederick, MD 21702, USA
1 To whom correspondence should be addressed: e-mail: gildersleevej{at}ncifcrf.gov
We recently reported carbohydrate microarray profile results for a set of commercially available antiglycan monoclonal antibodies (Manimala et al. 2007
), including clone A70-C/C8 (Kuemmel et al. 2007), which we acquired from Cell Sciences (Canton, MA), and antibodies B386 and B389, which we acquired from Biomeda (Foster City, CA). It was our understanding that B386 was equivalent to A68-B/A11 (Karsten et al. 1995) and B389 was equivalent to A70-A/A9 (Christensen et al. 2007). Subsequent to publication, we obtained A70-C/C8, A68-B/A11, and A70-A/A9 from the original source, Glycotope GmbH, Berlin-Buch (Dr. Uwe Karsten). In a side-by-side comparison, two of the antibodies supplied by Glycotope produced significantly different results as compared to the antibodies from Cell Sciences and Biomeda. In our assay, antibody A70-C/C8 from Glycotope bound selectively to LeY while A70-C/C8 from Cell Sciences cross-reacted with blood group B as we had reported. Antibody A68-B/A11 from Glycotope displayed reactivity with TF
, GA1 (TFβ), blood group A, blood group B, blood group H1, GM1, and tri-LacNAc while B386 from Biomeda bound blood group B had minor reactivity with blood group A trisaccharide and did not bind TF, GA1 (TFβ), blood group H1, GM1, or tri-LacNAc. Antibody A70-A/A9 from Glycotope and B389 from Biomeda both bound selectively to LeY and did not show appreciable differences. Based on these results, binding data, tissue staining, and Western blots obtained with a particular antibody should be interpreted in relation to the supplier. Table I lists the original sources and catalog numbers for the antibodies profiled in our previous paper (Manimala et al. 2007).
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References
Christensen PA, Danielczyk A, Ravn P, Stahn R, Karsten U, Goletz S. A monoclonal antibody to Lewis Y/Lewis b revealing mimicry of the histone H1 to carbohydrate structures. Scand J Immunol (2007) 65:362–367.[CrossRef][Web of Science][Medline]
Karsten U, Butschak G, Cao Y, Goletz S, Hanisch FG. A new monoclonal antibody (A78-G/A7) to the Thomsen–Friedenreich pan-tumor antigen. Hybridoma (1995) 14:37–44.[Web of Science][Medline]
Kuemmel A, Single K, Bittinger F, Faldum A, Schmidt LH, Sebastian M, Taube C, Buhl R, Wiewrodt R. The prognostic impact of blood group-related antigen Lewis Y and the ABH blood groups in resected non-small cell cancer. Tumor Biol (2007) 28:340–349.[CrossRef]
Manimala JC, Roach TA, Li Z, Gildersleeve JC. High-throughput carbohydrate microarray profiling of 27 antibodies demonstrates widespread specificity problems. Glycobiology (2007) 17:17C–23C.
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