Carrier hapten conjugation

The following is a generalized protocol for the activation of a protein with sulfo-SMCC with subsequent conjugation to a sulfhydryl-containing second molecule or protein. Specific examples of the use of this crosslinker to make antibody-enzyme or hapten-carrier conjugates may be found in Chapter 20, Section 1.1 and Chapter 19, Section 5, respectively. 

This chapter describes the design, preparation, and use of hapten-carrier conjugates used to elicit an immune response toward a coupled hapten. The chemical reactions discussed for these conjugations are useful for coupling peptides, proteins, carbohydrates, oligonucleotides, and other small organic molecules to various carrier macromolecules. The resultant conjugates are important in antibody production, immune response research, and in the creation of vaccines. 

Figure 19.13 Conjugation to KLH often can cause precipitation due to the high-molecular weight of the carrier protein. The conjugation of [Met5]-enkephalin to KLH yields a soluble immunogen if the level of EDC addition is about 0.1 times that typically used with BSA as a carrier. This figure shows the gel filtration pattern on Sephadex G-25 after completion of the crosslinking reaction. The first peak is KLH and the second peak is excess peptide. Depletion of the peptide peak correlates to hapten-carrier conjugation.

Purify the hapten-carrier conjugate by gel filtration or dialysis. 

NHS Ester-Maleimide Heterobifunctional Crosslinker-Mediated Hapten-Carrier Conjugation... 

The hapten-carrier conjugate now may be mixed with adjuvant and used for injection purposes without further purification. 

Diazo Cross-linked Hapten-Carrier Conjugate... 

Pedersen, M.K., Sorensen, N.S., Heegaard, P.M., Beyer, N.H., and Bruun, L. (2006) Effect of different hapten-carrier conjugation ratios and molecular orientations on antibody affinity against a peptide antigen./. Immunol. Meth. 311(1-2), 198-206. 

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