Determination of fibrinogen content in cryoprecipitated antihemophilic factor by fluorescence quenching method
Journal Title: Chinese Journal of Blood Transfusion - Year 2022, Vol 35, Issue 12
Abstract
Objective To establish a new method for the determination of fibrinogen content in cryoprecipitated antihemophilic factor. Methods Fibrinogen (Fib) could bind with sheep anti-human fibrinogen (anti-Fib) specifically and further form antigen-antibody complex. When the Fib was present in the solution, the fluorescence of fluorescein isothiocyanate (FITC) labeled on the anti-Fib (FITC-anti-Fib) was quenched due to the formation of immune complex. The fluorescence quenching degree of FITC-anti-Fib was positively correlated with Fib concentration (cFib) in a certain concentration range. Results The linear relationship between fluorescence quenching degree [(I0-I)/I0] of FITC-anti-Fib and ln(cFib) was (I0-I)/I0=15.53ln(cFib)+ 80.79 (R2=0.99) when the cFib was in the range of (0.007 8-0.560 0) g/L. The recovery of Fib was (96.77-102.43) %. When the method was applied to determine Fib at high, medium, and low concentrations, the obtained intra-day variation coefficients were 0.31%, 0.56%, and 0.49%, respectively, and the inter-day variation coefficients were 3.81%, 3.06%, and 4.13%, respectively. There was no significant difference between the results measured by fluorescence quenching method and coagulation method (t=-0.075, P>0.05). Conclusion In this work, a new fluorescence method for the determination of Fib in cryoprecipitated antihemophilic factor was successfully established based on the specific combination of fib and FITC-anti-Fib. The method is simple and rapid. The obtained results were accurate and reliable by using this method to determine Fib.
Authors and Affiliations
Jing LAN, Na FENG, Wenjuan ZHANG, Jin DING, Peng PENG, Na LIU, Xin CAO, Hua XU
Keywords
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- EP ID EP738428
- DOI 10.13303/j.cjbt.issn.1004-549x.2022.12.005
- Views 29
- Downloads 0
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