Monoclonal antibodies for the identification and purification of vNAR domains and IgNAR immunoglobulins from the horn shark Heterodontus francisci.

Karla Juarez; Gudrun Dubberke; Pavel Lugo; Friedrich Koch Nolte; Friedrich Buck; Friedrich Haag; Alexei Licea

Monoclonal antibodies for the identification and purification of vNAR domains and IgNAR immunoglobulins from the horn shark Heterodontus francisci.

Standard

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, Vol. 30, No. 4, 4, 2011, p. 323-329.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Juarez, K, Dubberke, G, Lugo, P, Koch Nolte, F, Buck, F, Haag, F & Licea, A 2011, 'Monoclonal antibodies for the identification and purification of vNAR domains and IgNAR immunoglobulins from the horn shark Heterodontus francisci.', HYBRIDOMA, vol. 30, no. 4, 4, pp. 323-329. <http://www.ncbi.nlm.nih.gov/pubmed/21851231?dopt=Citation>

APA

Juarez, K., Dubberke, G., Lugo, P., Koch Nolte, F., Buck, F., Haag, F., & Licea, A. (2011). Monoclonal antibodies for the identification and purification of vNAR domains and IgNAR immunoglobulins from the horn shark Heterodontus francisci. HYBRIDOMA, 30(4), 323-329. [4]. http://www.ncbi.nlm.nih.gov/pubmed/21851231?dopt=Citation

Vancouver

Juarez K, Dubberke G, Lugo P, Koch Nolte F, Buck F, Haag F et al. Monoclonal antibodies for the identification and purification of vNAR domains and IgNAR immunoglobulins from the horn shark Heterodontus francisci. HYBRIDOMA. 2011;30(4):323-329. 4.

Bibtex

@article{6aea5e90a2884603954c6505bda0f63c,

title = "Monoclonal antibodies for the identification and purification of vNAR domains and IgNAR immunoglobulins from the horn shark Heterodontus francisci.",

abstract = "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.",

keywords = "Animals, Humans, Cells, Cultured, Mice, Mice, Inbred BALB C, Amino Acid Sequence, Molecular Sequence Data, Immunoprecipitation, Protein Binding, Antibodies, Monoclonal, Murine-Derived/*genetics/immunology/metabolism, Antibody Specificity, Epitopes/chemistry, Erythrocytes/immunology, Fish Proteins/*genetics/immunology/metabolism, Hybridomas/metabolism, Immunity, Humoral, Immunoglobulin Variable Region/*genetics/immunology/metabolism, Immunoglobulins/genetics/*isolation & purification/metabolism, Peptide Fragments/chemistry, Recombinant Proteins/genetics/immunology/metabolism, Sharks/blood/*immunology, Animals, Humans, Cells, Cultured, Mice, Mice, Inbred BALB C, Amino Acid Sequence, Molecular Sequence Data, Immunoprecipitation, Protein Binding, Antibodies, Monoclonal, Murine-Derived/*genetics/immunology/metabolism, Antibody Specificity, Epitopes/chemistry, Erythrocytes/immunology, Fish Proteins/*genetics/immunology/metabolism, Hybridomas/metabolism, Immunity, Humoral, Immunoglobulin Variable Region/*genetics/immunology/metabolism, Immunoglobulins/genetics/*isolation & purification/metabolism, Peptide Fragments/chemistry, Recombinant Proteins/genetics/immunology/metabolism, Sharks/blood/*immunology",

author = "Karla Juarez and Gudrun Dubberke and Pavel Lugo and {Koch Nolte}, Friedrich and Friedrich Buck and Friedrich Haag and Alexei Licea",

year = "2011",

language = "English",

volume = "30",

pages = "323--329",

number = "4",

}

RIS

TY - JOUR

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