Validation of a fibrinogen γ’ enzyme-linked immunosorbent assay using a new monoclonal antibody: effects of bariatric surgery

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Abstract

Background: Fibrinogen γ’ is a naturally occurring 20-amino-acid splice variant of the fibrinogen γ chain. Animal studies link variations in fibrinogen to obesity, but it is unknown how fibrinogen γ’ is associated with obesity in humans. Objectives: To develop and validate an enzyme-linked immunosorbent assay (ELISA) for fibrinogen γ’ quantification in human plasma and analyze fibrinogen γ’ before and after bariatric surgery. Methods: We generated C-terminal fibrinogen γ’ specific mouse monoclonal antibodies and developed a γ’ ELISA. Validation included measures of accuracy, sensitivity, and precision. Fibrinogen γ’ and total fibrinogen were measured in 60 individuals before and 6 months after bariatric surgery and in 19 normal-weight controls and 120 blood donors. Results: Highly specific fibrinogen γ’ monoclonal antibodies were produced and successfully used in the ELISA. Recovery was 88%, and limits of detection and quantification were 0.003 mg/mL and 0.014 mg/mL, respectively. Coefficients of variation were 3% for repeatability and 7% for within-laboratory variation. The fibrinogen γ’ reference interval was 0.25 to 0.80 mg/mL. Fibrinogen γ’ concentrations were reduced after bariatric surgery and were higher in individuals with obesity than in those with normal weight. The fibrinogen γ’/total fibrinogen ratio was unchanged after surgery but was higher than the ratio in normal-weight individuals. Conclusion: We developed a precise and sensitive ELISA for fibrinogen γ’. Levels of fibrinogen γ’, but not the fibrinogen γ’/fibrinogen ratio, were reduced 6 months after bariatric surgery. Absolute and relative levels of fibrinogen γ’ were increased in individuals with obesity compared to normal-weight individuals.

Original languageEnglish
Article number102319
JournalResearch and Practice in Thrombosis and Haemostasis
Volume8
Issue number1
Number of pages10
ISSN2475-0379
DOIs
Publication statusPublished - Jan 2024

Bibliographical note

Publisher Copyright:
© 2024 The Authors

Keywords

  • antibodies, monoclonal
  • bariatric surgery
  • enzyme-linked immunosorbent assay
  • fibrinogen
  • obesity

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