- Email: [email protected]
Characterization of detergent extracts of human sperm membrane autoantigens by sera containing sperm autoantibodies
P 31
53
A NEW t ~ O D FOR ESTIMATION OF SPERM ANTIBODY TILERS INDIVIDUALLY TErnS OF ZgA, ZgG AND ZgM.
IN
A. B. CZUPPON, B. A. GE~J.~I, D. MAAS biotec diagnostica intl., Sterility Outpatient Clinic Medical School D-3000 Hannover I, F.R.G. The clinical significance of the antibody classes of spermatozoal antibodies in various specimens from patients with fertility problems has recently been recognized with respect to the choice of a suitable therapy (i). Up till now the various antibody classes of spermatozoal antibodies could only be determined by extremely complicated methods which could be performed in special laboratories only (i.e. chromatographic separation, density gradient ultracentrifugation etc. ) With the method presented here it is possible to check for IgA,IgG and IgM sperm antibody classes in little more then 60 min. Estimation of the individual titers is also possible. The new method can be used routinely in Sterility Clinics. Lit.:l) Z. Ulcova-Gallova et.al. Int.J.Fertil.,33,421-425
(1988)
Key~ords:Sperm antibody titer-lgA-IgG-IgM
P 32 CHARACTERIZATION OF DETERGENT EXTRACTS OF HUMAN SPERRRERBRIM~E AUTOANTIGENS BY SERACONTAINING SPERM AUTOANT]]~OIES
R.CHAWLA, S.P. AHUJA, S.SINGH, Department of Biochemistry, Punjab Agric.University, Ludhiana, India Human spermatozoal proteins (SMP) were extracted with 1,0 % ionic and nonionic detergent. Maximum amount of SMP were extracted by NP-40 and cetyi trimethyI ammonium bromide (CTAB). The carbohydrates were more in SMP extracted by deoxychoiate (DOC), CTAB and SDS, indicating giycoprotein (GP) nature of the same. These SMP were fractionated by PAGE and slo immunoreacted with WHO sara # CDS 65,67,95,97,98,101 and Bank serum # 101 having sperm autoantibodies (SAAb). The immune compiexes (SIC) thus formed couid onIy be precipitated with 7.5 % poiyethyiene giycoi (PEG, moi.wt. 6 Kd). Maximum SIC were precipitated by PEG from CTAB extract reacted with sere # CDS 67,97,98,101. AII the sera except # 65 formed SIC with DOC extract (in iesser amounts). None of the sera detected autoantigens (AAg) in Triton X-tO0 and Brij-35 extracts. Sera # 95 and 97 formed SIC with NP-40 extract. The PAGE pattern of SIC showed that CTAB, DOC and NP-40 extracted GP-AAg (high and moderate moi.wt: RM 0.1 and 0.4 - 0.5). However, NP-40 extract had an additionai GP-AAg (Iow moi.wt. RM 0.73). The resuits suggested that the SM AAg extracted by CTAB, DOC and NP-40 can be purified by reacting with sperm autoantisera foiIowed by precipitation with 7.5 % PEG.
Human sperm membrane autoantigens characterization
53
A NEW t ~ O D FOR ESTIMATION OF SPERM ANTIBODY TILERS INDIVIDUALLY TErnS OF ZgA, ZgG AND ZgM.
IN
A. B. CZUPPON, B. A. GE~J.~I, D. MAAS biotec diagnostica intl., Sterility Outpatient Clinic Medical School D-3000 Hannover I, F.R.G. The clinical significance of the antibody classes of spermatozoal antibodies in various specimens from patients with fertility problems has recently been recognized with respect to the choice of a suitable therapy (i). Up till now the various antibody classes of spermatozoal antibodies could only be determined by extremely complicated methods which could be performed in special laboratories only (i.e. chromatographic separation, density gradient ultracentrifugation etc. ) With the method presented here it is possible to check for IgA,IgG and IgM sperm antibody classes in little more then 60 min. Estimation of the individual titers is also possible. The new method can be used routinely in Sterility Clinics. Lit.:l) Z. Ulcova-Gallova et.al. Int.J.Fertil.,33,421-425
(1988)
Key~ords:Sperm antibody titer-lgA-IgG-IgM
P 32 CHARACTERIZATION OF DETERGENT EXTRACTS OF HUMAN SPERRRERBRIM~E AUTOANTIGENS BY SERACONTAINING SPERM AUTOANT]]~OIES
R.CHAWLA, S.P. AHUJA, S.SINGH, Department of Biochemistry, Punjab Agric.University, Ludhiana, India Human spermatozoal proteins (SMP) were extracted with 1,0 % ionic and nonionic detergent. Maximum amount of SMP were extracted by NP-40 and cetyi trimethyI ammonium bromide (CTAB). The carbohydrates were more in SMP extracted by deoxychoiate (DOC), CTAB and SDS, indicating giycoprotein (GP) nature of the same. These SMP were fractionated by PAGE and slo immunoreacted with WHO sara # CDS 65,67,95,97,98,101 and Bank serum # 101 having sperm autoantibodies (SAAb). The immune compiexes (SIC) thus formed couid onIy be precipitated with 7.5 % poiyethyiene giycoi (PEG, moi.wt. 6 Kd). Maximum SIC were precipitated by PEG from CTAB extract reacted with sere # CDS 67,97,98,101. AII the sera except # 65 formed SIC with DOC extract (in iesser amounts). None of the sera detected autoantigens (AAg) in Triton X-tO0 and Brij-35 extracts. Sera # 95 and 97 formed SIC with NP-40 extract. The PAGE pattern of SIC showed that CTAB, DOC and NP-40 extracted GP-AAg (high and moderate moi.wt: RM 0.1 and 0.4 - 0.5). However, NP-40 extract had an additionai GP-AAg (Iow moi.wt. RM 0.73). The resuits suggested that the SM AAg extracted by CTAB, DOC and NP-40 can be purified by reacting with sperm autoantisera foiIowed by precipitation with 7.5 % PEG.
Human sperm membrane autoantigens characterization