Bioengineering and modification of antibody as a capture molecule is very important to increase sensitivity and specificity of the biosensor. As a detecting molecule to diagnose Legionella infectious disease, PAL (peptidoglycanassociated lipoprotein) specific antibody was immunoaffinity purified from rabbit polyclonal anti-PAL IgG by immobilizing PAL onto CNBr-activated sepharose. For the immunoassay, intact antibody was modified into F(ab’)2 and either intact antibody (H+L) or F(ab’)2 was layered on carboxyl terminated PAMAM dendrimer (generation 4.5), which was immobilized onto an amine functionalized polystyrene surface. Subsequently, soluble antigen of Legionella pneumophila serogroup 1 or Legionella anisa diluted with sodium phosphate buffer or patient’s plasma was applied respectively and detected by HRPconjugated immunoaffinity purified rabbit polyclonal anti-PAL. As a result, each antibody on dendrimer presented better binding activity than the antibody on blank surface. Moreover, F(ab’)2 on dendrimer showed higher binding capacity than intact antibody on dendrimer against soluble Legionella antigens diluted in plasma. This study demonstrated modified antibody immobilized on dendrimer amplified the signal by increasing binding capacity/sensitivity and minimizing steric hindrance. Furthermore, this sensor format demonstrated it could be applied as a protein biosensor in clinical samples. (The authors gratefully acknowledge supplying materials from the laboratory of Dr. Minja Kim, Division of Infectious Disease, Department of Internal Medicine, College of Medicine, Korea University.)
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
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