A Sensitive Monoclonal Antibody Sandwich ELISA for the Measurement of the Major Olive Pollen Allergen Ole e 1

van Ree, Ronald; Aalbers, Marja; Kea, Olga; Marco de la Calle, Francisco M.; Sempere Ortells, José M.; Villalba, Mayte; Rodriguez, Rosalia; Aalberse, Rob C.
July 2000
International Archives of Allergy & Immunology;2000, Vol. 122 Issue 3, p224
Academic Journal
Background: Olive pollen is a major cause of inhalant allergy in countries around the Mediterranean sea. The major allergen of olive pollen is Ole e 1. Measurement of the major allergen content of allergen products for diagnosis and therapy is becoming an essential element of standardization protocols. This study aimed at the development of a monoclonal antibody (mAb) sandwich ELISA for Ole e 1. Methods: Balb/c mice immunized with Ole e 1 were used for the production of mAbs. Screening of mice and hybridomas was performed in a RIA with radiolabeled purified Ole e 1. Purified mAbs were used as catching and/or (biotinylated) detecting antibodies in sandwich ELISA. Results: Four mAbs (IgG1κ) directed to nonoverlapping epitopes on Ole e 1 were obtained: 1A12, 5C1, 10A12 and 3H8. Both 1A12 and 10A12 were successfully used for affinity purification of Ole e 1 from olive pollen extract. Two sandwich ELISAs were developed, with 1A12 and 10A12 as catching, and 5C1 and 3H8 as detecting antibodies, respectively. Both catching and detecting antibodies were used in similar concentrations, ranging from 60 to 100 ng/well. For both ELISAs, the sensitivity was approximately 1 ng/ml of Ole e 1. The measuring range was from 1 to 25 ng/ml. No significant differences were observed, when the performance of both ELISAs in standardization of olive pollen extracts was compared. Conclusions: Two sensitive sandwich ELISAs for the major olive pollen allergen Ole e 1 were developed. They will prove to be useful tools in allergen standardization protocols.Copyright © 2000 S. Karger AG, Basel


Related Articles

  • Humanization of an Anti-CD34 Monoclonal Antibody by Complementarity-determining Region Grafting Based on Computer-assisted Molecular Modelling. Sheng Hou; Bohua Li; Ling Wang; Weizhu Qian; Dapeng Zhang; Xueyu Hong; Hao Wang; Yajun Guo // Journal of Biochemistry;Jul2008, Vol. 144 Issue 1, p115 

    4C8 is a new mouse anti-human CD34 monoclonal antibody (mAb), which recognizes class II CD34 epitopes and can be used for clinical hematopoietic stem/progenitor cell selection. In an attempt to improve its safety profiles, we have developed a humanized antibody of 4C8 by...

  • Successful in vivo tumor targeting of prostate-specific membrane antigen with a highly efficient J591/PEI/DNA molecular conjugate. Moffatt, S.; Papasakelariou, C.; Wiehle, S.; Cristiano, R. // Gene Therapy;May2006, Vol. 13 Issue 9, p761 

    We have utilized a novel polyethylenimine (PEI)/DNA-βgal vector to investigate the specificity and efficiency of immuno-targeting prostate-specific membrane antigen (PSMA). Coupling of the PSMA-specific monoclonal antibody, J591, to the vector was facilitated via the high-affinity interaction...

  • Limitation of microwave treatment for double immunolabelling with antibodies of the same species and isotype. Bauer, Mario; Schilling, Nicole; Spanel-Borowski, Katharina // Histochemistry & Cell Biology;Sep2001, Vol. 116 Issue 3, p227 

    Microwave treatment (MW) involves completely blocking contaminating staining in the double-labelling technique, using primary monoclonal antibodies from the same species and the same isotype as well as the same secondary antibody (ab). However, we noticed some limitations when locating...

  • RT1.L: a family of MHC class Ib genes of the rat major histocompatibility complex with a distinct promoter structure. Lambracht-Washington, Doris; Düvel, Heike; Hänisch, Lisa; Dinkel, Astrid; Wonigeit, Kurt // Immunogenetics;Apr2004, Vol. 56 Issue 1, p28 

    RT1.L class I antigens have originally been identified in LEW rats by LEW.1LV3-anti-LEW.1LM1 antisera and have been classified as nonclassical. We report now that LEW.1LV3-anti-LEW.1LM1 antisera react with three different antigens, termed RT1.L1, RT1.L2, and RT1.L3. This was found by serological...

  • Very late antigen-5 and leukocyte function-associated antigen-1 are critical for early stage hematopoietic progenitor cell homing. Asaumi, N.; Omoto, E.; Mahmut, N.; Katayama, Y.; Takeda, K.; Shinagawa, K.; Harada, M. // Annals of Hematology;Jul2001, Vol. 80 Issue 7, p387 

    Interactions of adhesion molecules among hematopoietic progenitor cells (HPC), bone marrow microvascular endothelial cells (BMMEC), and stromal cells are critical for hematopoiesis. However, most of the identified HPC receptors mediate interactions between HPC and stromal cells in the...

  • monoclonal antibody.  // Taber's Cyclopedic Medical Dictionary (2009);2009, Issue 21, p1486 

    An encyclopedia entry for "monoclonal antibody," which refers to the antibody which is specific to a certain antigen, is presented.

  • Monoclonal Antibodies against Blo t 13, a Recombinant Allergen from Blomia tropicalis. Labrada, Mayrel; Uyema, Keiko; Sewer, Minerva; Labrada, Alexis; González, Maritza; Caraballo, Luis; Puerta, Leonardo // International Archives of Allergy & Immunology;2002, Vol. 129 Issue 3, p212 

    Background: The recombinant allergen of Blomia tropicalis, rBlo t 13, shows 11% of IgE reactivity to sera from allergic patients. This allergen belongs to the fatty acid-binding protein family and its natural equivalent remains to be isolated. Monoclonal antibodies (MAbs) are important tools for...

  • Immunoglobulin: DNA cofactor interaction. Malinka, M.; Petriev, V.; Podgorodnichenko, V. // Pharmaceutical Chemistry Journal;Jun2007, Vol. 41 Issue 6, p289 

    The reasons for the appearance of DNA-binding activity of immunoglobulins (Ig) after chromatography on anion-exchange Sephadexes were studied. Elution of Ig from these Sephadexes with 0.5 M NaCl was found, using an orcinol method, to be associated with degradation of the matrix, with the...

  • Immunoradiometric assay for the rapid detection of HLA-B27. Trapani, Joseph A.; Vaughan, Hilary A.; Tait, Brian D.; Mckenzie, Ian F. C. // Immunology & Cell Biology;Jun1988, Vol. 66 Issue 3, p215 

    A new method for the rapid detection of the HLA-B27 antigen is discussed which consists of a direct immunoradiometric assay (IRA) utilizing a 125I-labelled. HLA-B27 specific monoclonal antibody to detect HLA-B27 in whole citrated blood. Thus far, the assay has been used to assign HLA-B27 status...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics