Production of Polyclonal Antibody Against Recombinant Growth Hormone and Designing an Elisa Kit and Comparing Some of Its Diagnostics Indices With a Commercial Kit



Maneian M^{1, 6} ; Zarkesh Esfahani SH^{2, 3} ; Akbari M⁴ ; Khanahmad H⁵ ; Masjedi M^{1, 6}
Source: Journal of Isfahan Medical School Published:2013

Abstract

Background: Human growth hormone (hGH) is a secretory protein of the blood which is used extensively in biotechnology and medical sciences. Recombinant human growth hormone production is still one of the priorities of the Iranian Ministry of Health. One of the most sensitive and accurate techniques for the measurement of this hormone is the ELISA method. Scientists need to produce antibody against hGH to measure the serum level of hGH in patients. Commercial ELISA kits are commonly imported from other countries to Iran. Therefore, the current study aimed to produce a polyclonal antibody against hGH in order to make a domestic ELISA kit to measure this hormone. Methods: Rabbit polyclonal antibody against hGH was produced following 10 weeks of immunization by the antigen injection. The antibody characteristics were evaluated by immunoblotting. The purified antibody was used to design the ELISA kit. The designed sandwich ELISA method was standardized concerning the amount of antibody coating and antibody conjugate. Ultimately, the sensitivity and specificity of the designed kit was compared with the foreign kits. Findings: The polyclonal antibody against hGH was produced in rabbits and purified successfully. This antibody can be used in the western blot and ELISA methods. The diagnostic value of the designed kit was 98%, compared with the foreign kits. Conclusion: This kit has a diagnostic value in the assessment of hGH; therefore, it can be a good alternative to the commercial kits. Therefore, this antibody and ELISA kit can be produced more cost-effectively and can be a step towards self-sufficiency. © 2013, Isfahan University of Medical Sciences(IUMS). All rights reserved.