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Antibodies

Range of Antibody Specifications and Applications

The various specificities offered for research include blood group related, extracellular (such as Band 3, GPA and GPC), intracellular (such as Band 3, GPA, GPC and Glut-1) and cytoskeletal antibodies (such as ankyrin, spectrin and Protein 4.2). There are also monoclonal antibodies with specificities to platelets, granulocytes and leucocytes, HLA complement, NK and secondary reagents.

Many of the monoclonal antibodies are specific for the following CD markers:

The CE marked Feto-Maternal Haemorrhage (FMH) reagents include both BRAD 3 FITC (anti-D) and AEVZ 5.3 FITC (isotype matched negative control). FITC conjugated BRAD 3 quantitates the numbers of D positive cells in a mixture of D positve and negative cells, allowing estimation of the size of feto-maternal bleeds in D negative mothers with a D positive fetus. An additional reagent, BIRMA 17C PE, has specificity for CD66b (granulocytes) and complements the method for FMH estimation using a two-colour method which allows the elimination (by gating) of interfering neutrophils, thereby ensuring a more accurate measurement.

Purification of Antibodies

Antibodies are supplied as purified immunoglobulin. 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 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 and so the methods have been optimised for each antibody. Purity of antibodies is checked by non-reducing SDS polyacrylamide gel electrophoresis, ELISA, serological methods and flow cytometry. Purified antibodies are stored at high concentration in aliquots at -80°C. QC tests are carried out as for the supernatant batches.

Conjugated Antibodies

Purified antibodies are supplied conjugated to either FITC, R-phycoerythrin (R-PE), Allophycocyanin (APC), HRP or Biotin. Conjugation to other fluorochromes can be requested on an individual basis. Several QC tests are carried out to ensure that the conjugated products are active and specific.

Culture supernatants, purified antibodies and conjugated antibodies can be produced on a large scale and can be sold on a collaborative or commercial basis.

Two FITC conjugated antibodies and one PE conjugated antibody have been CE marked.

We have the capacity and skills to produce both Fab and F(ab)2 fragments. This is only carried out on an individual basis following communications with the customer. Specific requests can be made about product specifications, which must be agreed with PDPU before work can be undertaken.

Discounts may be available to customers seeking to purchase in bulk quantitites.



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