Subramanian Sarada, Karandeb Anjali A, Adiga P Radhakantha
Department of Neurochemistry, National Institute of Mental Health and Neurosciences, Bangalore 560 029, India.
Indian J Biochem Biophys. 2004 Dec;41(6):281-7.
Chicken riboflavin carrier protein (RCP; 219 AA) harbours four linear epitopes, constituted by the peptide residues 3-23, 64-83, 130-147 and 200-219. Antibodies to these sequences bioneutralize maternal RCP and provide protection from pregnancy in rodents. In order to overcome the major histocompatibility complex-dependent variability in immune response often encountered with use of single peptides for vaccination in genetically outbred populations, we have assembled a novel synthetic gene, incorporating in tandem the nucleotide sequences coding for all the four neutralizing epitopes of chicken RCP and expressed in Escherichia coli. The gene product, mini-RCP has been characterized for its immunogenic properties and contraceptive potential in rodents. Immunization of rabbits and rats led to generation of antibodies against individual peptide components, as determined by enzyme-linked-immunosorbent assay (ELISA). However, immunized rats carried pregnancy to term and delivered healthy offsprings. Antisera from these rats exhibited decreased affinity of binding to the native protein. These findings suggest that the prospects of covalently-linked epitope peptides need to be cautiously evaluated during the design and development of peptide-based vaccines.
鸡核黄素载体蛋白(RCP;219个氨基酸)含有四个线性表位,由肽残基3 - 23、64 - 83、130 - 147和200 - 219构成。针对这些序列的抗体可生物中和母体RCP,并在啮齿动物中提供避孕保护。为了克服在遗传上远交群体中使用单一肽进行疫苗接种时经常遇到的主要组织相容性复合体依赖性免疫反应变异性,我们组装了一个新的合成基因,串联整合编码鸡RCP所有四个中和表位的核苷酸序列,并在大肠杆菌中表达。该基因产物mini - RCP已在啮齿动物中对其免疫原性和避孕潜力进行了表征。通过酶联免疫吸附测定(ELISA)确定,对兔和大鼠进行免疫导致产生针对各个肽成分的抗体。然而,免疫后的大鼠足月妊娠并产下健康后代。这些大鼠的抗血清与天然蛋白的结合亲和力降低。这些发现表明,在基于肽的疫苗的设计和开发过程中,需要谨慎评估共价连接的表位肽的前景。