Monoclonal antibodies for the identification and purification of vNAR domains and IgNAR immunoglobulins from the horn shark Heterodontus francisci.
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Monoclonal antibodies for the identification and purification of vNAR domains and IgNAR immunoglobulins from the horn shark Heterodontus francisci. / Juarez, Karla; Dubberke, Gudrun; Lugo, Pavel; Koch Nolte, Friedrich; Buck, Friedrich; Haag, Friedrich; Licea, Alexei.
in: HYBRIDOMA, Jahrgang 30, Nr. 4, 4, 2011, S. 323-329.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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T1 - Monoclonal antibodies for the identification and purification of vNAR domains and IgNAR immunoglobulins from the horn shark Heterodontus francisci.
AU - Juarez, Karla
AU - Dubberke, Gudrun
AU - Lugo, Pavel
AU - Koch Nolte, Friedrich
AU - Buck, Friedrich
AU - Haag, Friedrich
AU - Licea, Alexei
PY - 2011
Y1 - 2011
N2 - In addition to conventional antibodies, cartilaginous fish have evolved a distinctive type of immunoglobulin, designated as IgNAR, which lacks the light polypeptide chains and is composed entirely by heavy chains. IgNAR molecules can be manipulated by molecular engineering to produce the variable domain of a single heavy chain polypeptide (vNARs). These, together with the VHH camel domains, constitute the smallest naturally occurring domains able to recognize an antigen. Their special features, such as small size, long extended finger-like CDR3, and thermal and chemical stability, make them suitable candidates for biotechnological purposes. Here we describe the generation of two mouse monoclonal antibodies (MAbs), MAb 370-12 and MAb 533-10, that both specifically react with vNAR domains of the horn shark Heterodontus francisci. While the former recognizes a broad spectrum of recombinant vNAR proteins, the latter is more restricted. MAb 370-12 precipitated a single band from whole shark serum, which was identified as IgNAR by mass spectrometry. Additionally, we used MAb 370-12 to follow the IgNAR-mediated immune response of sharks during immunization protocols with two different antigens (complete cells and a synthethic peptide), thus corroborating that MAb 370-12 recognizes both isolated vNAR domains and whole IgNAR molecules. Both MAbs represent an affordable molecular, biochemical, and biotechnological tool in the field of shark single-domain antibodies.
AB - In addition to conventional antibodies, cartilaginous fish have evolved a distinctive type of immunoglobulin, designated as IgNAR, which lacks the light polypeptide chains and is composed entirely by heavy chains. IgNAR molecules can be manipulated by molecular engineering to produce the variable domain of a single heavy chain polypeptide (vNARs). These, together with the VHH camel domains, constitute the smallest naturally occurring domains able to recognize an antigen. Their special features, such as small size, long extended finger-like CDR3, and thermal and chemical stability, make them suitable candidates for biotechnological purposes. Here we describe the generation of two mouse monoclonal antibodies (MAbs), MAb 370-12 and MAb 533-10, that both specifically react with vNAR domains of the horn shark Heterodontus francisci. While the former recognizes a broad spectrum of recombinant vNAR proteins, the latter is more restricted. MAb 370-12 precipitated a single band from whole shark serum, which was identified as IgNAR by mass spectrometry. Additionally, we used MAb 370-12 to follow the IgNAR-mediated immune response of sharks during immunization protocols with two different antigens (complete cells and a synthethic peptide), thus corroborating that MAb 370-12 recognizes both isolated vNAR domains and whole IgNAR molecules. Both MAbs represent an affordable molecular, biochemical, and biotechnological tool in the field of shark single-domain antibodies.
KW - Animals
KW - Humans
KW - Cells, Cultured
KW - Mice
KW - Mice, Inbred BALB C
KW - Amino Acid Sequence
KW - Molecular Sequence Data
KW - Immunoprecipitation
KW - Protein Binding
KW - Antibodies, Monoclonal, Murine-Derived/genetics/immunology/metabolism
KW - Antibody Specificity
KW - Epitopes/chemistry
KW - Erythrocytes/immunology
KW - Fish Proteins/genetics/immunology/metabolism
KW - Hybridomas/metabolism
KW - Immunity, Humoral
KW - Immunoglobulin Variable Region/genetics/immunology/metabolism
KW - Immunoglobulins/genetics/isolation & purification/metabolism
KW - Peptide Fragments/chemistry
KW - Recombinant Proteins/genetics/immunology/metabolism
KW - Sharks/blood/immunology
KW - Animals
KW - Humans
KW - Cells, Cultured
KW - Mice
KW - Mice, Inbred BALB C
KW - Amino Acid Sequence
KW - Molecular Sequence Data
KW - Immunoprecipitation
KW - Protein Binding
KW - Antibodies, Monoclonal, Murine-Derived/genetics/immunology/metabolism
KW - Antibody Specificity
KW - Epitopes/chemistry
KW - Erythrocytes/immunology
KW - Fish Proteins/genetics/immunology/metabolism
KW - Hybridomas/metabolism
KW - Immunity, Humoral
KW - Immunoglobulin Variable Region/genetics/immunology/metabolism
KW - Immunoglobulins/genetics/isolation & purification/metabolism
KW - Peptide Fragments/chemistry
KW - Recombinant Proteins/genetics/immunology/metabolism
KW - Sharks/blood/immunology
M3 - SCORING: Journal article
VL - 30
SP - 323
EP - 329
IS - 4
M1 - 4
ER -