An improved phage-display panning method to produce an HM-1 killer toxin anti-idiotypic antibody

Kabir, M. Enamul; Krishnaswamy, Senthilkumar; Miyamoto, Masahiko; Furuichi, Yasuhiro; Komiyama, Tomoyoshi

doi:10.1186/1472-6750-9-99

Cited by 22 publications

(12 citation statements)

References 42 publications

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“…Various endeavours have been taken to select good binders from antibody phage libraries against various antigens. Approaches such as increased library sizes [96,100], enhanced cloning strategies [153], and improve panning protocols [154] have been demonstrated to retrieve good binders. Another approach reported the use of protease sensitive helper phage KM13.…”

Section: Naïve Library Derived Antibodies Against Target Antigens Of mentioning

confidence: 99%

Naïve Human Antibody Libraries for Infectious Diseases

Chan

Rahumatullah

Lai

et al. 2017

Advances in Experimental Medicine and Biology

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Many countries are facing an uphill battle in combating the spread of infectious diseases. The constant evolution of microorganisms magnifies the problem as it facilitates the re-emergence of old infectious diseases as well as promote the introduction of new and more deadly variants. Evidently, infectious diseases have contributed to an alarming rate of mortality worldwide making it a growing concern. Historically, antibodies have been used successfully to prevent and treat infectious diseases since the nineteenth century using antisera collected from immunized animals. The inherent ability of antibodies to trigger effector mechanisms aids the immune system to fight off pathogens that invades the host. Immune libraries have always been an important source of antibodies for infectious diseases due to the skewed repertoire generated post infection. Even so, the role and ability of naïve antibody libraries should not be underestimated. The naïve repertoire has its own unique advantages in generating antibodies against target antigens. This chapter will highlight the concept, advantages and application of human naïve libraries as a source to isolate antibodies against infectious disease target antigens.

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Section: Naïve Library Derived Antibodies Against Target Antigens Of mentioning

confidence: 99%

Naïve Human Antibody Libraries for Infectious Diseases

Chan

Rahumatullah

Lai

et al. 2017

Advances in Experimental Medicine and Biology

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“…However, these drugs have repeatedly failed against infections caused by Cryptococcus species (Kordossis et al, 1998;Saag et al, 2000). Therefore, the development of novel molecules and alternative therapeutic strategies for the battle against cryptococcosis as well as other opportunistic fungal infections is becoming a topical and widely recognized need (Devi, 1996;Kordossis et al, 1998;Larsen et al, 2005;Kabir et al, 2009). Monoclonal antibody therapy is a rational alternative for the treatment of Crytococcus neoformans infections in immunocompromized patients resulting from AIDS, transplant recipients, and cancer chemotherapy.…”

Section: Introductionmentioning

confidence: 98%

“…Taking advantage of its immunegenicity and a potent ß-1,3-glucan synthase inhibitor, HM-1 killer toxin neutralizing monoclonal antibody (nmAb-KT) has been produced. To produce a new antifungal drug, we used nmAb-KT as an immunogen to generate recombinant anti-idotypic scFv antibodies that share structural and functional similarities with the active site of HM-1 (Selvakumar et al, 2006b;Kabir et al, 2009;Krishnaswamy et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

New avenues for phage‐display library to produce a Cryptococcus‐specific anti‐idiotypic antibody of HM‐1 killer toxin

Rahman

Kabir

Krishnaswamy

et al. 2011

J of Molecular Recognition

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Existing antifungal drugs are notable for their inability to act rapidly, as well as their toxicity and limited spectrum. The identification of fungal-specific genes and virulence factors would provide targets for new and influential drugs. The display of repertories of antibody fragments on the surface of filamentous phage offers a new way to produce immunoreagents as defined specificities. Here we report the selection of Cryptococcus-specific targets by using phage-display panning from a cDNA library, where bactericidal antibodies have been developed against conserved surface-exposed antigens. A single-chain variable fragment (scFv) phage library was constructed from splenocyte of an immunized mouse by idiotypic vaccination with HM-1 killer toxin (HM-1) neutralizing monoclonal antibody (nmAb-KT) that was used for selection against Cryptococcus neoformans membrane fraction (CnMF). Key elements were the selection against antigen (nmAb-KT and CnMF) and the release of bound phages using competitive panning elution with CnMF at neutral pH condition. Isolated scFvs react specifically with C. neoformans and some other pathogenic and non-pathogenic fungal strain's cell wall receptors by exerting strong antifungal activity in vitro. A high affinity clone, designated M1 was selected for detailed characterization and tested anti-cryptococcal activity with IC(50) values at 5.33 × 10(-7) to 5.56 × 10(-7) M against C. neoformans. The method described here is a new technique for the isolation of cell membrane specific immunoreactive phages in the form of scFv using CnMF that contained cell membrane associated proteins.

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“…To overcome the intrinsic toxicity and chemical instability of HM‐1 and simultaneously take advantage of its immunogenicity, nmAb‐KT has been produced . With the aim of producing a new anti‐fungal drug, we used nmAb‐KT as an immunogen in mice to produce an antigen‐specific anti‐idiotypic antibody that shares structural and/or functional similarities with the active site of HM‐1 .…”

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confidence: 99%

Antigen itself antibody: an alternative one‐step immunoassay for measuring the anti‐idiotypic antibody titer

Kabir

Krishnaswamy

Selvakumar

et al. 2014

Microbiology and Immunology

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Immunoassay designs rely on the great specificity of antibodies and a suitable marker that facilitates generation of a quantitative signal. Currently, there is no reliable method for measuring the titers of an anti-idiotypic antibody. Our initial attempt to measure titers of mouse anti-idiotypic antibody after idiotypic vaccination with HM-1 killer toxin neutralizing monoclonal antibody (nmAb-KT) failed. Because the injected antigen, nmAb-KT, is a mouse IgG, using a commercial antibody to measure the antibody titer always gave a false positive signal against control mouse serum antibody in parallel with the antigen-treated immunized serum antibodies. To get a reliable and clearly differentiable signal by ELISA, idiotypic antigen was labeled with HRP and HRP-conjugated-nmAb-KT used to measure the antibody titers in the antigen-treated mice. Compared with control mice, signals were found in high anti-nmAb-KT IgG responses in test mice; however, untreated control mice had a significant amount of purified non-specific IgG. This method is amenable to long read lengths and will likely enable anti-idiotypic antibody titer measurement in a more specific and cost effective way without requiring commercial antibody.Key words anti-idiotypic antibody, killer toxin neutralizing monoclonal antibody, one-step immunoassay, titer measurement.Killer toxins are protein molecules that disrupt cell functions in a number of ways (1, 2). Among the yeast killer toxins, HM-1 is highly stable against heat treatment and a wide range of pH values (pH 2-11); it also exhibits a wide spectrum of anti-fungal activity (3, 4). These unique properties have made this toxin attractive for therapeutic applications, but not for clinical use both because of its instability in host physiological conditions and its antigenicity (2, 4, 5). To overcome the intrinsic toxicity and chemical instability of HM-1 and simultaneously take advantage of its immunogenicity, nmAb-KT has been produced (6). With the aim of producing a new anti-fungal drug, we used nmAb-KT as an immunogen in mice to produce an antigenspecific anti-idiotypic antibody that shares structural and/or functional similarities with the active site of HM-1 (7-15).Antibodies are highly specific, naturally evolved molecules that recognize specific antigens and can be classified according to their mode of action as they react to and set about defending the body against foreign invaders (16). Anti-idiotypic antibodies are a special set

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An improved phage-display panning method to produce an HM-1 killer toxin anti-idiotypic antibody

Cited by 22 publications

References 42 publications

Naïve Human Antibody Libraries for Infectious Diseases

Naïve Human Antibody Libraries for Infectious Diseases

New avenues for phage‐display library to produce a Cryptococcus‐specific anti‐idiotypic antibody of HM‐1 killer toxin

Antigen itself antibody: an alternative one‐step immunoassay for measuring the anti‐idiotypic antibody titer

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