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001-es BibID:	BIBFORM043016
035-os BibID:	PMID:22797718
Első szerző:	Shrestha, Dilip (biológus)
Cím:	Comparative study of the three different fluorophore antibody conjugation strategies / Dilip Shrestha, Adrienn Bagosi, János Szöllősi, Attila Jenei
Dátum:	2012
Megjegyzések:	The progression in bioconjugational chemistry has significantly contributed to the evolution and success of protein biology. Mainly, antibody chemistry has been a subject of intensive study owing to the expansion of research areas warranted by using various derivatives of conjugated antibodies. Three reactive moieties (amine, sulfhydryl and carbohydrate) in the antibodies are chiefly favored for the conjugational purpose. This feature is known for decades, nevertheless, amine based conjugation is still the most preferred strategy despite the appreciation the other two methods receive in conserving the antigen binding affinity (ABA). No single report has been published, according to our knowledge, where these three conjugation strategies were applied to the same fluorophore antibody systems. In this study, we evaluated conjugation yield, time demand and cost efficiency of these conjugation procedures. Our results showed that amine based conjugations was by far the best technique due to its simplicity, rapidity, ease of operation, higher conjugate yield, cheaper cost and potential for larger fluorophore/protein labeling ratio without having much effect in ABA. Furthermore, sulfhydryl labeling clearly excelled in terms of reduced non-specific binding and mild effect in ABA but was usually complicated by an asymmetric antibody reduction due to mercaptoethylamine while carbohydrate oxidation based strategy performed the worst during our experiment
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban Antibodies Antibody conjugation article binding affinity Biophysics cell biology chemistry Comparative Study Hungary methods Research Research Support Support time egyetemen (Magyarországon) készült közlemény
Megjelenés:	Analytical and Bioanalytical Chemistry - 404 : 5 (2012), p. 1449-1463. -
További szerzők:	Bagosi Adrienn Szöllősi János (1953-) (biofizikus) Jenei Attila (1966-) (biofizikus)
Pályázati támogatás:	TÁMOP-4.2.2-08/1-2008-0019 TÁMOP TÁMOP-4.2.1/B-09/1/KONV-2010-007 TÁMOP TÁMOP-4.2.2/B-10/ 1-2010-0024 TÁMOP
Internet cím:	DOI Intézményi repozitóriumban (DEA) tárolt változat
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