Department of Neurology and Translational Research Center, Rigshospitalet, Glostrup, Denmark.
Department of Autoimmunology and Biomarkers, Statens Serum Institut, Copenhagen, Denmark.
Methods Mol Biol. 2024;2821:65-70. doi: 10.1007/978-1-0716-3914-6_5.
Conjugation to carrier proteins is necessary for peptides to be able to induce antibody formation when injected into animals together with a suitable adjuvant. This is usually performed by conjugation in solution followed by mixing with the adjuvant. Alternatively, the carrier may be adsorbed onto a solid support followed by activation and conjugation with the peptide by solid-phase chemistry. Different reagents can be used for conjugation through peptide functional groups (-SH, -NH2, -COOH), and various carrier proteins may be used depending on the peptides and the intended use of the antibodies. The solid phase may be an ion exchange matrix, from which the conjugate can subsequently be eluted and mixed with adjuvant. Alternatively, the adjuvant aluminum hydroxide may be used as the solid-phase matrix, whereupon the carrier is immobilized and conjugated with peptide. The resulting adjuvant-carrier-peptide complexes may then be used directly for immunization.
与合适的佐剂一起注射到动物体内时,载体蛋白的缀合对于肽能够诱导抗体形成是必要的。这通常通过溶液中的缀合来完成,然后与佐剂混合。或者,载体可以吸附到固体载体上,然后通过固相化学与肽进行活化和缀合。可以使用不同的试剂通过肽的官能团(-SH、-NH2、-COOH)进行缀合,并且根据肽和抗体的预期用途,可以使用各种载体蛋白。固相可以是离子交换基质,随后可以从该基质中洗脱缀合物并与佐剂混合。或者,可以使用佐剂氢氧化铝作为固相基质,载体在此固定化并与肽缀合。然后可以直接使用所得的佐剂-载体-肽复合物进行免疫。