Insulin Aspart ELISA

Insulin aspart is a fast-acting insulin analog marketed by Novo Nordisk as NovoLog/NovoRapid. It is a manufactured form of human insulin; where a single amino acid has been exchanged. This change helps the fast-acting insulin analog be absorbed quickly into the bloodstream. As a result, it starts working in minutes, which allows one to take insulin and eat right away. Fast-acting insulin analogs are considered to act similarly to the way insulin is released in people without diabetes mellitus.

It is a recombinant, biosynthetic, fast-acting insulin analogue. Compared to human insulin, it has a single amino acid substitution at position B28 where proline is replaced with aspartic acid. This substitution decreases its propensity to form hexamers and gives it a higher rate of absorption following subcutaneous administration compared to native insulin.

Principle of the assay: The method employs the quantitative sandwich enzyme immunoassay technique. In the first step, samples and standards are pipetted into microwells. If Insulin Aspart i s present in samples or standards, it will form complex with Anti-Insulin Aspart antibody, which is pre-coated onto microwells. In the second step, Anti-Insulin Aspart-Biotin conjugate is pipetted and incubated. In the third step, HRP Conjugate is pipetted and incubated. Free HRP conjugate will be removed by a washing step. In the fourth step , TM B substrate is added to microwells and color develops proportionally to the amount of Insulin Aspart present in samples or standards. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.