Poh Chit Laa, Yahaya Abdul Aziz Al-Fattah Bin, Cheong Huey Tyng, Lim Hui Xuan
Centre for Virus and Vaccine Research, School of Medical and Life Sciences, Sunway University, Selangor, Malaysia.
Methods Mol Biol. 2024;2821:165-177. doi: 10.1007/978-1-0716-3914-6_13.
Vaccination is an effective means of inducing immune protection to prevent transmissible diseases. During the Covid-19 pandemic, immunizations using traditional and novel vaccine platforms such as the inactivated SARSCo-V-2 vaccine, adenoviral-vectored, and nucleic acid-based mRNA vaccines have been relatively successful in controlling the rates of infection and hospitalizations. Nevertheless, the danger posed by the emergence of SARS-CoV-2 variants would set the stage for the design of next generation vaccines. To overcome the lack of efficacy of current vaccines against emerging SARS-CoV-2 variants, new vaccines must be able to overcome the reduced effectiveness of the current vaccines. Since the current Covid-19 vaccines are dependent on the whole S-protein of Wuhan strain as the antigen, mutations have rendered the current Covid-19 vaccines less effective against variants of concern (VoCs). Instead of using the whole S-protein, peptide-based epitopes could be predicted using immunoinformatic approaches, simulation of the 3D structures, overlapping peptides covering the whole length of the S-protein or peptide arrays based on synthetic peptide combinatorial libraries comprising peptides recognizable by monoclonal antibodies. B-cell epitopes were predicted, and immunogenicity of peptides was validated in mice by immunizing mice with peptides conjugated to keyhole limpet hemocyanin (KLH) mixed with Montanide 51 as an adjuvant. The immunogenicity of epitopes that could elicit peptide specific IgGs was determined by peptide-based ELISA. Neutralizing activities were determined by cPass and pseudovirus-based neutralization assays.
疫苗接种是诱导免疫保护以预防传染病的有效手段。在新冠疫情期间,使用传统和新型疫苗平台(如灭活的严重急性呼吸综合征冠状病毒2(SARS-CoV-2)疫苗、腺病毒载体疫苗和基于核酸的信使核糖核酸(mRNA)疫苗)进行免疫接种在控制感染率和住院率方面相对成功。然而,SARS-CoV-2变体的出现所带来的危险将为下一代疫苗的设计奠定基础。为了克服当前疫苗对新出现的SARS-CoV-2变体缺乏效力的问题,新疫苗必须能够克服当前疫苗效力降低的问题。由于当前的新冠疫苗依赖武汉株的全长刺突蛋白(S蛋白)作为抗原,突变使得当前的新冠疫苗对关注变体(VoC)的效力降低。可以使用免疫信息学方法、3D结构模拟、覆盖S蛋白全长的重叠肽或基于包含可被单克隆抗体识别的肽的合成肽组合文库的肽阵列来预测基于肽的表位,而不是使用全长S蛋白。预测了B细胞表位,并通过用与钥孔血蓝蛋白(KLH)偶联的肽免疫小鼠(将其与Montanide 51作为佐剂混合)在小鼠中验证了肽的免疫原性。通过基于肽的酶联免疫吸附测定(ELISA)确定可引发肽特异性免疫球蛋白G(IgG)的表位的免疫原性。通过cPass和基于假病毒的中和试验确定中和活性。