I etertnarv Immunology and Immunopathologv 37 ( 1993 ) 61-71 61
Else, ler Science Pubhshers B V, Amsterdam
Purification and characterization of IgG and
IgG2 from buffalo (Bubalus bubalis) serum and
colostrum
N K Kakker and M C Goel
lmmunologl Se~ tton Department o! ~ etermat )' Ml~ robtolog~ Hat vana 4gpt~ ultmal (~m) et sttl
Htsar-125004 lndta
( Accepted 10 August 1992)
ABSTRACT
Kakker N K and Goel M C, 1993 Purification and characterization oflgG~ and IgG2 from buffalo
(Bubalus bubah~ ) serum and colostrum Iet hnmunol hnmunopathol 37 61-71
Buffalo lgG~ and IgG2 were purified from serum and colostrum using salt preclp,tatlon, dial)sis
gel filtration and ion-exchange chromatograph) Their purity was monitored b) lmmunodfffusLon
and mlmunoelectrophoresis using antl-heav} chain specific sera and SDS-PAGE Selectl~ e binding ol
IgG2 to protem-A was used to remo~e lgG2 from lgGl preparations The IgGt and IgG2 had a molec-
ular mass (Mr) of 162 0 and 161 5 kD, respectively and were found to consist o! hea~) (H) and light
( L ) chains The H and L chains had Mr of 58 and 24 kD respect~ el~ Reductmn-alk~latlon followed
by gel filtration was used for the ~solatlon of H and k chains While intact H chains were obtained the
L chains appeared to be cleaved into 14 kD molecules and smaller fragments The mean hexoscs
content of the serum lgG~ and IgGz was 1 81 ± 0 02% and 0 70 _+ 0 02% respectl~el~ The correspond-
mg ~alues for colostral lgG~ and IgG~ were 1 76+0 01% and 0 78_+0 08% Both the IgG subclasses
activated homologous complement These results suggest that buffalo and cattle IgG subclasses ha~ e
man) common characteristics and minor differences
ABBREVIATIONS
C, complement, EA erythrocyte-antlbody, H, heavy, HA, haemagglutmation ID, ~mmunod~ffu-
slon IE lmmunoelectrophoresls Igs lmmunoglobuhns L light 2-ME 2-mercaptoethanol
INTRODUCTION
The Immunoglobuhns (Igs) of cattle have been extensively investigated
and reviewed (Butler, 1983, 1986), but not those of the water buffalo, an
economically important species Buffaloes hke other ruminants, possess at
least three classes of Igs, namely IgM, IgA and IgG with ~ts two subclasses,
Correspondence to N K Kakker, Immunology Section, Department of Veterlnar~ M~croblol-
og) Haryana Agricultural University, Hlsar-125004, India
© 1993 Elsevier Science Publishers B V