免疫
免疫原性
基因枪
裸DNA
生物
单克隆抗体
噬菌体展示
抗体
病毒学
免疫系统
免疫学
计算生物学
基因
遗传增强
dna疫苗
遗传学
作者
Frédéric Trempe,Martín A. Rossotti,Tahir Maqbool,C. Roger MacKenzie,Mehdi Arbabi‐Ghahroudi
标识
DOI:10.1007/978-1-0716-2075-5_3
摘要
Genetic immunization is a simple, cost-effective, and powerful tool for inducing innate and adaptive immune responses to combat infectious diseases and difficult-to-treat illnesses. DNA immunization is increasingly used in the generation of monoclonal antibodies against targets for which pure proteins are unavailable or are difficult to express and purify (e.g., ion channels and receptors, transmembrane proteins, and emerging infectious pathogens). Genetic immunization has been successfully utilized in small inbred laboratory animals (mostly rodents); however, low immunogenicity of DNA/RNA injected into large mammals, including humans, is still a major challenge. Here, we provide a method for the genetic immunization of llamas, using a combination of biolistic transfection with a gene gun and intradermal injection with a DERMOJET® device, to elicit heavy-chain IgG responses against epidermal growth factor receptor (EGFR). We show the technique can be used to generate single-domain antibodies (VHHs) with nanomolar affinities to EGFR. We provide methods for gene gun bullet preparation, llama immunization, serology, phage-display library construction and panning, and VHH characterization.
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