[Comparison of polyclonal anti-sera against extracellular domain of hepatoma associated antigen HAb18G/CD147 prepared by different immunization schemes].
OBJECTIVE
To compare characteristics of polyclonal anti-sera against extracellular domain of hepatoma-associated antigen HAb18G/CD147(HAb18GEF) generated by different immunization schemes.


METHODS
BALB/c mice were immunized with GST-HAb18GEF fusion protein expressed in E.coli (routine immunization method), recombinant eukaryotic expression plasmid pcDNA3/HAb18G (intramuscular injection) and pcDNA3/HAb18G plasmid followed by human hepatoma cells booster (DNA-cell booster), respectively. The titers and Ig subclasses of polyclonal anti-sera against denatured and natural HAb18GEF were detected by indirect ELISA and cell ELISA, respectively. The specific binding of polyclonal anti-sera prepared by different immunization schemes to denatured HAb18GEF was analyzed by Western blot. The specific binding of polyclonal anti-sera produced by DNA-cell booster immunization to natural HAb18G antigen on hepatoma cells was detected by immunofluorescence staining.


RESULTS
GST-HAb18GEF immunization could induce polycolonal antibody IgG1 with higher titer mainly against denatured or linear epitopes on HAb18GEF. Antibody induced by pcDNA3/HAb18G intramuscular immunization was IgG2a with lower titer against natural epitopes on HAb18GEF. DNA-cell booster immunization could induce generation of polycolonal antibody IgG2a and IgG1 of moderate titers against the native epitopes on HAb18G.


CONCLUSIONS
The polycolonal sera with different titers against different epitopes on HAb18GEF can be induced by different immunization schemes.
Zhang
Si-He
SH
Cell Engineering Research Center, Fourth Military Medical University, Xi'an 710032, China.
Yang
Xiang-Min
XM
Xing
Jin-Liang
JL
Chen
Zhi-Nan
ZN
D000818:Animals; D000918:Antibody Specificity; D051926:Basigin / Q000737:chemistry* / Q000276:immunology*; D006528:Carcinoma, Hepatocellular / Q000276:immunology* / Q000473:pathology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005110:Extracellular Space / Q000276:immunology*; D006801:Humans; D007106:Immune Sera / Q000276:immunology*; D007114:Immunization; D051379:Mice; D017434:Protein Structure, Tertiary
D000939:Epitopes; D007106:Immune Sera; D051926:Basigin