No chromatography required Solulink’s AP-Antibody All-in-One Conjugation Kit offers an innovative TurboLink™ catalyzed conjugation based on Solulink’s proprietary linking technology that delivers 100% conversion of antibody to conjugate that generates a conjugate free of unlabeled antibody or enzyme at a high final yield of > 40%, withoutHPLC.The kit includes everything needed to make two direct-labeled AP-antibody conjugates from 100 µg of any user-supplied antibody.
S-HyNic (succinimidyl-6-hydrazino-nicotinamide) linker, an NHS-ester, reacts with lysine residues, incorporating HyNic functional groups (hydrazinonicotinamide) onto the antibody.
Sulfo-S-4FB (succinimidyl-4-formylbenzamide) linker is conjugated to AP, providing a pre-activated, high-activity AP called 4FB-AP.
HyNic-modified antibody is incubated with pre-activated 4FB-AP in the presence of the TurboLink™ catalyst, aniline, leading to rapid and efficient conversion of the antibody to conjugate through formation of stable bis-arylhydrazone bonds.
Catalyzed conjugation The efficiency of catalyzed conjugation greatly simplifies conjugate purification. Solulink’s proprietary linking technology is catalyzed by the TurboLink™ catalyst, aniline, under mild reaction conditions, increasing the rate and efficiency, leading to quantitative conversion of free antibody to conjugate. The complete absence of free antibody at the end of the catalyzed reaction leaves only two components; excess AP and conjugate. Purified conjugate is then eluted from the filter membrane, free of residual antibody and AP in high yield (40–60 μg).