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editorial
. 2008 Jun 6;18(10):746. doi: 10.1093/glycob/cwn049

Supplier-dependent antiglycan monoclonal antibody specificities: Comment on “High-throughput carbohydrate microarray profiling of 27 antibodies demonstrates widespread specificity problems”

Jeff Gildersleeve 2,1, Timothy A Roach 2, Zhitao Li 2, Jeffrey C Gildersleeve 2
PMCID: PMC2733772  PMID: 18539626

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 TFα, 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 TFα, 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).

Table I.

Sources and catalog numbers for the antibodies

Clone Listed antigen Source company Catalog number
81FR2.2 BG-A Dako Cytomation A0581
B480 BG-A Biomeda V3251
CLCP-19B BG-B EMD 202769
B460 BG-B Biomeda V7071
B389 BG-H2, Ley, Leb Biomeda V7025
92FR-A2 BG-H2 Biomeda V1087
B376 BG-H2 Biomeda V7023
A63-D/B12 Ley, BG-H2 Glycotope MAB-P205
B393 BG-H2 Biomeda V7022
A46-B/B10 Ley, BG-H2 Glycotope MAB-P203
7LE Lea Lab Vision MS-494 – P0
PR.5C5 Lea Chemicon CBL205
T174 Lea EMD 434627
121SLE SLea Lab Vision MS-379 – P0
CA199.02 SLea Lab Vision MS-1166 – P0
2-25LE Leb Lab Vision MS-1024 – P0
T218 Leb EMD 434629
K21 Lec Genetex GTX23352
15C02 Lex Lab Vision MS-739 – S0
28 Lex Chemicon CBL144
ZC-18C Lex Chemicon MAB1205
FR4A5 Lex Lab Vision MS-430 – P1
BR55 Ley Glycotech 11-006
F3 Ley EMD 434636
A70-C/C8 Ley Cell Sciences MON2047
B1.1 Tn Biomeda V1053
B386 TF Biomeda V7028
1A4 Gb3 Glycotech 11–008
5B5 Gb3 BD Biosciences 551352

References

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