Developing rapid test kits for detection of heart attack and congestive heart failure

The aim of this thesis was to obtain hybridoma cells that produce anti-hFABP and anti-NT-proBNP monoclonal antibodies to design sensitive diagnostic test kits for detectioon of heart attack and congestive heart failure. For this purpose, hybridoma clones that produce anti-hFABP and anti-NT-proBNP monoclonal antibodies were obtained by in vitro immunization methodology and characterized by growth kinetics, isotyping and HPLC. Lateral flow immonuassays (LFIA) and localized-surface plasmon resonance (LSPR)-based diagnostic test kits in the form of 96 well plate and microfluidic chips were designed by using produced antibodies in combination of commercial ones. The detection limits of the lateral flow immonuassays (LFIA) were determined as 10 ng/mL and 1000 ng/mL for hFABP and NT-proBNP in PBS, respectively. The sensitivity of the LSPR-based diagnostic kits in both 96 well plate and microfluidic chip forms were found to be dramatically increased compared to that of LFIA. 96 well plate LSPR-based diagnostic kits was found to be able to detect 1 pg/mL hFABP in PBS and 10 pg/ml hFABP in serum while detecting 100 pg/mL NT-proBNP in PBS and 1 pg/mL NT-proBNP in serum. The sensitivity of the single-use microfluidic chips was determined as 10 pg/ml for hFABP and NT-proBNP in PBS. Consequently, LFIA based assays are easy to produce, portable, requires no reading system with less sensitivity. On the other hand, LSPR based systems are far more sensitive than developped LFIA kits (10 pg/mL). The only disadvantage of LSPR-based kits is the need of a spectrophotometer for analysis. But with the production of portable LSPR-based microfluidic chips this disadvantage is not going to be concern with high sensitive, easy to use portable set-ups.