HCID 178476261, compound → HCID 178476260, compound

Participants

Reaction Steps

Six mg of the GnRH-Gly-Cys hapten (containing an free SH on one end) is dissolved in 1 ml of H2O. The dissolved hapten is added to the activated KLH and allowed to react for 2 hours at room temperature and then overnight in the refrigerator. The KLH-hapten conjugate is immediately purified by applying the reaction mixture to a desalting column. (Sephadex G-25). The conjugate protein should come off on the void volume (first peak, fractions 4-6) as measure by absorbance at 280 nm. There should be a drop in absorbance after fraction 6 and rise in absorbance in fractions 7 or 8 as the excess hapten comes off. A small amount of excess hapten does not cause problem in a prime dose but could neutralize antibody in a boost dose if it is present in large excess. The GnRH/KLH conjugate may be frozen or lyophilized or used in the present form.

Provenance & Evidence