Description
Introduction:
The KRIBIOLISA ELISA kits are used for assessing the specific biomarker in samples analytes which may be serum and cell culture supernatant as validated with the kit. The kit employs a blocking ELISA technique which engages the blocking pathway to estimate the neutralizing antibodies. Imiglucerase is a form of recombinant human beta-glucocerebrosidase enzyme used to replace the deficient endogenous enzyme in the treatment of Gaucher disease. Human Beta-glucocerebrosidase or Beta-D-glucosyl-N-acylsphingosine glucohydrolase E.C. 3.2.1.45. 497 residue protein with N-linked carbohydrates, MW=59.3 kD. Alglucerase is prepared by modification of the oligosaccharide chains of human Beta-glucocerebrosidase. The modification alters the sugar residues at the non-reducing ends of the oligosaccharide chains of the glycoprotein so that they are predominantly terminated with mannose residues. A neutralizing antibody (NAb) is an antibody that is responsible for defending cells from pathogens and are produced naturally by the body as part of its immune response. Their production is triggered by both infections and vaccinations against infections. In an immunogenetic context it will bind to a drug and neutralize its therapeutic effect.
Intended Use:
The KRIBIOLISA Neutralizing Antibodies to Imiglucerase (Cerezyme) ELISA kit is used as an analytical tool for the qualitative detection of neutralizing antibodies against Imiglucerase in serum or plasma.
Principle:
The method employs sandwich ELISA technique. The protein-protein interaction between HRP-Imiglucerase and Anti-Imiglucerase can be blocked by Imiglucerase. The neutralizing antibodies against Imiglucerase bind to Imiglucerase and thus a competitive inhibition is created. Samples and controls are pipetted in a blank microtitre plate and incubated with neutralizing antibody to Imiglucerase (NAb) and Imiglucerase. This complex of bound and unbound Imiglucerase is then incubated with HRP conjugated Imiglucerase protein. This complex solution of bound antibodies to Imiglucerase and unbound Imiglucerase is then pipetted into Anti Imiglucerase coated microplate. After washing, the substrate solution (TMB) is added to the microwells. Post incubation, color develops proportionate to the amount of bound HRP:Imiglucerase. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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