Preliminary Study Of Latex-enhanced Immunoturbidimetric Assay Of Rh-endostatin And Transferrin Detection Kit

Endostatin is known as one of tumor angiogenesis inhibitor, and can be used for the treatment of solid tumors. Clinically the detection of endostatin concentration is by ELISA method. But this method is time-consuming and expensive, so we hope to be able to find other effctive method which can rapidly and conveniently detect endostatin. Immuno latex turbidimetric method is operated automated, repeated well, can reduce the nonspecific agglutination and has a low false positive rate. We therefore self-made latex-enhanced immunoturbidimetric assay of rh-endostatin detection kit by marking the appropriate size of latex with rh-endostatin antibody, taking good linear range and larger range as the standard for reagen components and testing conditions to detect the rh-endostatin standard by single factor experiment.The results show that latex with the same size of 130 nm and EDAC, NHS and rh-endostatin can covalently bound and form sensitized latex. The optimum testing condition is at 0.3% concentration of sensitized latex,510 nm wavelength,2.5% concentration of PEG 6000,8μL for the sample amount, NaN₃ of preservative and reacting for 10 min after adding the sensitized latex.Transferrin has a prominent role in antibacterial, bactericidal and tumors and cancer prevention. In order to be able to detect transferrin concentration much more easily, we hope to develp latex-enhanced immunoturbidimetric assay of transferrin detection kit. In this study, the transferrin gene containing plasmid was selected as a template to amplify the target gene by PCR. The target fragment and linear plasmids were botained from the transferrin gene and the expression vector pET28a with dual digestion using Xho I and Nde I, respectively. The expression vector pET28a containing trasferrin gene was built using T4 DNA ligase, and provides conditions for the expression and purification of transferrin, for the reparation of transferrin antibody and for the self-made latex-enhanced immunoturbidimetric assay of transferrin detection kit.