Elimination of interfering activity in serum samples in the Chinese hamster ovary pertussis serology assay.
An interfering substance in various frozen serum samples was observed to inhibit the adhesion of Chinese hamster ovary (CHO) cells to microplate surfaces during a CHO pertussis neutralization test, resulting in wells that lacked cells or wells with dead cells after 2 days of incubation. The interfering activity in the serum could be eliminated by (i) transferring cells to other wells after their initial incubation, (ii) adding fetal calf serum (FCS) to the sample dilution buffer, (iii) precoating microplates with FCS, or (iv) preincubating the samples at 4 degrees C for 5 days. Preincubating the samples at 4 degrees C for 5 days reduced the interfering activity in only some of the samples. Adding serum to the sample dilution buffer or precoating the microplates with serum did not influence the antibody titers in the serum samples. The method described may be used for routine applications.
Østergaard
Erik
E
Quality Control Department, Statens Serum Institut, 5 Artillerivej, DK-2300 Copenhagen S, Denmark.
Sørensen
Charlotte
C
Nielsen
Lene
L
Stawski
Gitte
G
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media; D005260:Female; D006801:Humans; D009500:Neutralization Tests; D037342:Pertussis Toxin / Q000276:immunology*; D010567:Pertussis Vaccine / Q000276:immunology*; D012699:Serology / Q000379:methods*; D044967:Serum / Q000737:chemistry*
D000907:Antibodies, Bacterial; D003470:Culture Media; D010567:Pertussis Vaccine; D037342:Pertussis Toxin