Recombinant expression and partial charachterication of anti cardiac troponin ScFv in Pichia pastoris

Heart and vascular diseases are one of the major cause of all deaths around the world with 30% incidence. Therefore, detection of myocardial infarction has a great importance in responding to emergence cases for increasing the survival rate. Immune dependent quick diagnostic kits have a key role in this point. Anti-cTnI antibody is mostly produced in recombinant Chinese Hamster Ovary (CHO) cell lines and partly in host animals such as mouse and rabbit. However, heterogeneity of antibodies produced with these seras exhibit some disadvantageous difficulties for development of diagnostic test kits. In this study anti-cardiac troponin I (anti-cTnI) antibody, which is one of the best diagnostic antibodies used in myocardial infract diagnosis overall the world, was recombinantly expressed as an antibody fragment (hypervariable region) in Pichia pastoris at a concentration of 12.3 mg/l. Recombinant ScFv was purified using immobilized metal ion affinity chromatography. Its aplicability in these diagnostic kits before and after conjugation to gold nanoparticles was evaluated by enzyme linked immunosorbent assay. By utilization of Pichia expression system, it is intended to reduce variations (antibody heterogeneity) among whole antibodies. According to the literature, this is the first study where a recombinant anti-troponin I antibody fragment has been expressed to use in diagnostic kit development. The data obtained serve a guideway for a technology aiming to produce similar antibody fragments for diagnostic purposes.