Active immunization of beef heifers against luteinizing hormone: II. Evaluation of conjugation technique on antigenicity of LH.

This study evaluated the antigenicity of LH-ovalbumin complexes produced using different conjugation techniques. Two homobifunctional cross-linkers, glutaraldehyde (Glut) and carbodiimide (ECDI), were evaluated together with one heterobifunctional reagent, m-maleimido-benzoyl N-hydroxysuccinimide ester (MBS). Polyacrylamide gel electrophoresis and Western transfer techniques were used to confirm conjugation of LH. Forty-four beef heifers were assigned randomly to seven treatment groups. Two groups of heifers were immunized against glutaraldehyde conjugates (Glut-I and Glut-II), two against MBS conjugates (MBS-I and MBS-II) and one against a carbodiimide conjugate (ECDI). Control animals were immunized against nonconjugated LH (LH-only) or ovalbumin alone (Oval). Heifers received one primary injection of antigen followed by two boosters at a 3-wk interval. The Glut conjugates induced the highest (P less than .05) LH antibody activity (Glut-II, 18 +/- 4%; Glut-I, 14 +/- 4%). The ECDI (11 +/- 4%), and MBS-I (11 +/- 2%) conjugates induced greater LH binding than MBS-II (4 +/- 1%), LH-only (4 +/- 1%) or Oval (2 +/- 1%). Glutaraldehyde produced an LH-ovalbumin conjugate of greater LH immunogenicity than either ECDI or the heterobifunctional reagent, MBS.