Establishment of an OPK method for the detection of functional antibody in immune serum of Staphylococcus aureus vaccine
ZHOU Heng
YANG Liuyang
YANG Feng
ZHANG Jinyong
ZOU Jintao
CHENG Ping
ZENG Hao
ZOU Quanming
Abstract：The study is aimed to establish a method to detect functional antibodies in the immune serum of Staphylococcus aureus vaccine.According to the University of Albama at Birmingham Protocol for multiplexed opsonophagocytic killing assay (UAB-MOPA) method,we established the HL-60 effect cell cultures,S.aureus stocks,and selected the optimal complement concentration,serum pre-dilution fold and the incubation time of opsonization and phagocytosis phase.Then OPK assay was performed by using five different S.aureus strains.Data showed that after HL-60 cells were differentiated in 0.8％ DMF culture for 4 days,the expression of CD35 was 87.1％,the expression of CD71 was 2.82％,the percentage of living cells was 75.7％.Target bacteria work resurrection rate was ＞90％,while dilution degrees of bacterial stocks were ＞1 000;complement in the final concentration was 6％.The opsonization and phagocytosis phase times were confirmed as 30 minutes and 60 minutes.Serum was pre-diluted 2-folds,and vaccinated serum showed OPK activity against five different S.aureus strains,and all curves exhibited standard "S" shape.In conclusion,we established the OPK detection method for recombinant S.aureus subunit vaccine,which could effectively evaluate the serum functional antibody in vitro,thus provides an important technical support for the effective alternative destination of clinical trials.
Keywords：S.aureusRecombinant subunit vaccineFunctional antibodyOpsonnophagocytic killing assay
Publication Date：2018-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 252-258 )