Multivalent scFv Display of Phagemid Repertoires for the Selection of Carbohydrate-specific Antibodies and its Application to the Thomsen–Friedenreich Antigen

Highly Enhanced Cytotoxicity of a Dimeric Bispecific Diabody, the hEx3 Tetrabody

“…The multimerization of small antibody fragments can lead to improved pharmacokinetics and binding affinity, resulting in an enhancement of the therapeutic effect. To increase the population of hEx3 tetramers for therapeutic application, we are working to modify the middle linker in hetero-scFvs, to change the orientation of VH and VL, and to create mutations to minimize the steric interference in the tetrameric form, similar to how the scFv multimer has been modified (35,38,39).…”

Generation of antibodies recognizing an aberrant glycoform of human tissue inhibitor of metalloproteinase-1 (TIMP-1) using decoy immunization and phage display

“…The cDNAs encoding mouse heavy chain and kappa chain variable regions (VH and VK) were then amplified by PCR using Ex Taq polymerase (Takara, Shiga, Japan) and degenerate primers specific for VH and VK (Wang et al, 2000). For multivalent scFv phagemid library format based on shortened linker, single residue linker was designed to be introduced between VH and VK, as described by Ravn et al (Ravn et al, 2004). Briefly, the amplified VH and VK containing BbsI site were digested with BbsI and joined by ligation.…”

“…Nonetheless, immunization with carbohydrates is not always successful and often leads to a weak primary IgM response or no response in some instances because carbohydrates are self-antigens. Alternatively, phage-displayed antibody library has also been used to isolate anti-glycan antibodies Mao et al, 1999;Ravn et al, 2004;Sakai et al, 2007;Schoonbroodt et al, 2008). Phage display can give an artificial flexibility in antibody design such as valency and diversity, and is considered to be more advantageous for low immunogenic self-antigen like carbohydrates (Ravn et al, 2004;Schoonbroodt et al, 2008).…”

Purification and refolding of anti-T-antigen single chain antibodies (scFvs) expressed in Escherichia coli as inclusion bodies

“…In the authors' laboratory, phage display technology has been used to obtain scFv genes with specificities for carbohydrate moieties since many carbohydrates are self-antigens that seldom induce an immune response in animals (6,7). Genes encoding anti-Man3, anti-Lewis X,…”