AUTOIMMUNE SKIN DISEASE

Instructions for use

The MESACUP anti-Type VII collagen TEST is a semi-quantitative enzyme-linked immunosorbent assay(ELISA) for the detection of anti-type VII collagen antibodies in serum.

Background:

Epidermolysis bullosa aquisita (EBA) is an autoimmune blistering skin disease characterized by the presence of Type VII Collagen IgG autoantibodies. Type VII collagen is a major component of anchoring fibrils, attachment structures within the basement membrane, between the epidermis and dermis of human skin. Type VII collagen is composed of three identical α-chains. Each α-chain consists of a central collagenus (COL) domain flanked by a 145 kDa non-collagenous amino-terminal (NC1) domain, and a smaller 34 kDa carboxyl-terminal non-collagen (NC2) domain. The major antigenic epitopes for anti-Type VII collagen autoantibodies in patients with EBA reside within the NC1 domain (1-3). However, the recent research reports that NC2 domain is a minor antigenic epitopes (4). To develop an ELISA assay with the highest sensitivity to identify Type VII Collagen antibodies, two purified recombinant antigens (NC1 and NC2) were combined and coated in the same well of ELISA microplate.

The MESACUP Anti-Type collagen VII TEST measures anti-type VII collagen autoantibodies containing not only anti-NC1antibody but also anti-NC2 antibody in patient serum specifically.

Principle:

The MESACUP Anti-Type VII collagen TEST measures anti-Type VII collagen antibodies present in the serum by ELISA. Calibrators and patient sera are added to microwells coated with two kinds of recombinant human NC1 and NC2 antigen, allowing anti-Type VII collagen antibodies to react with the immobilized antigen (Sample incubation). After washing to remove any unbound serum proteins, horseradish peroxidase conjugated anti human IgG antibody is added and incubated (Conjugate incubation). Following another washing step, the peroxidase substrate is added and incubated for an additional period of time (Substrate incubation). Acid solution is then added to each well to terminate the enzyme reaction and to stabilize the color development. The assay can be quantified by measuring the reaction photometrically.

Size: 48 wells

Method: ELISA

Antigen: recombinant