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Range of Antibody Specifications and Applications

Examples of Use:

The anti-DAF (CD55) and CD59 antibodies are of interest to workers studying the regulation of complement activity in health and disease.
Anti-LFA-3 (CD58) and CD44 antibodies are useful in studies of the mechanisms of antigen specific immune recognition and lymphocyte migration.

BRAD 3 is a human monoclonal anti-D that reacts as an indirect agglutinin with all Rh D positive red cells tested except those of the rare DVI or Rohar types (pattern 135). By flow cytometry, FITC-BRAD 3 can be used to quantitate accurately the numbers of Rh D positive cells in a mixture of Rh D positive and negative cells, and thereby estimate the size of feto-maternal bleeds6,7. BRAD 3 can also discriminate weak D in feto-maternal bleeds where the site/cell numbers are above 1000 RhD sites8,9. FITC conjugated BRAD 3 has also been used in conjunction with R-phycoerythrin conjugated BRIC 256 (anti-Glycophorin A) in a dual labelling flow cytometry method for FMH quantitation10. BCSH Guidelines for FMH have been published11. When measuring the variance of rr cells in terms of background binding of FITC conjugated IgG, the use of a negative control FITC-labelled antibody should be used in parallel with FITC anti-D on clinical samples12. At the end of 2006, both BRAD 3 FITC and AEVZ 5.3 FITC negative control were CE marked as a kit to be used to measure FMH. In 2011, an agreement was made with IQ Products ( Netherlands ) to distribute this kit worldwide.

CD233 (Band 3). These antibodies react to different epitopes located on either the extracellular or intracellular domains. CD233 associates with the erythrocyte membrane protein glycophorin A (GPA) which promotes the correct folding and translocation during biosynthesis of CD233. Many CD233 mutations are known in man and these mutations can lead to two types of disease; destabilization of the erythrocyte membrane leading to hereditary spherocytosis, and defective kidney acid secretion leading to distal renal tubular acidosis. Other CD233 mutations that do not give rise to disease result in novel blood group antigens, which form the Diego blood group system.

IBGRL Research Products also have a range of antibodies that react with platelets (GPIa/IIa or CD29/CD49b, GPIIb/IIIa or CD41/CD61and CD42b), granulocytes (CD66b) and HLA. The range includes antibodies that react with NK cells and one that reacts with complement C3d.




Purification of Antibodies

Antibodies can be supplied as purified immunoglobulin on request, although most antibodies are available from stock. IgG antibodies are purified by affinity chromatography on a protein-G Sepharose column in an FPLC system. IgM antibodies are purified by a variety of different techniques including affinity chromatography (using an anti-mouse IgM column), ion exchange chromatography followed by size exclusion chromatography on Sephacryl S-300 or more recently using a capture select method. Unfortunately, not one single method can be applied for IgM purification as for each different IgM, the yields vary with each technique. The methods have to be optimised for each antibody. Purity of antibodies is checked by non-reducing SDS polyacrylamide gel electrophoresis, ELISA, serological methods or flow cytometry. Purified antibodies are stored at high concentration in aliquots at -80C. QC tests are carried out as for the supernatant batches.

Conjugated Antibodies

Purified antibodies conjugated to either FITC, R-phycoerythrin (R-PE) or Biotin have been made available since 1994. Other conjugations can be requested on an individual basis as we have conjugated antibodies to peroxidase and Alexa Fluor 488. Again, various QC tests are carried out to ensure that the products are active and specific.
Culture supernatants, purified antibodies and conjugated antibodies can be produced on a large scale according to GLP and can be sold on a collaborative or commercial basis. Two FITC conjugated antibodies have been CE marked. We have the capacity and skills to produce both Fab and F(ab)2 fragments but this is only carried out on an individual basis following communications with the customer. Specific requests can be made with regard to product specifications but this must be agreed with IBGRL before it is undertaken. We are happy to negotiate on price depending on the amount of product requested; please ask for a quote.