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BACKGROUND Grass pollen extracts are complex mixtures consisting of different major allergenic and non-allergenic components. Phl p 4 is an important allergen, because more than 75% of grass pollen allergic patients produce specific IgE antibodies against group 4 allergens. OBJECTIVE This study was designed to investigate the specificity of monoclonal antibodies (MoAbs) produced against Phl p 4 and to verify the presence of group 4-like proteins in different grass pollen. Furthermore the usefulness of MoAbs for quantification of group 4 allergens was studied. METHODS Group 4 analogues were investigated by immunoblotting and ELISA inhibition using three MoAbs. The specificity of antibodies was studied using isolated group 1 and group 5 allergens. Quantification of group 4 allergen was achieved by a two-site solid-phase ELISA. Phl p 4 was purified from whole pollen extract by chromatographic or electrophoretic techniques and used as standard. RESULTS The MoAbs studied bound strongly to proteins from timothy grass pollen extract at a mw of 55 kDa and a pI of 9.0-9.3. Phl p 4 homologes with similar mw were detected in Dactylis glomerata, Festuca pratensis, Holcus lanatus, Poa pratensis, Lolium perenne. Epitope mapping showed that all three MoAb recognized unrelated regions on Phl p 4. A two-site binding ELISA using MoAbs was developed for determination of Phl p 4 in Phleum pratense extracts. The method was able to evaluate group 4 in mass units with a working range between 150 and 2000 ng/mL. The absolute amounts of group 4 in extracts of several grasses varied considerably but was always-less than 1% of the total protein. CONCLUSIONS Group 4 homologes are present in the various grass extracts but to different extents. The group 4 ELISA could be very useful as a additional tool for providing information concerning the composition of grass pollen extracts.
B Fahlbusch, and
W D Müller, and
O Rudeschko, and
L Jäger, and
O Cromwell, and
H Fiebig
March 2004,
The Journal of allergy and clinical immunology,
B Fahlbusch, and
W D Müller, and
O Rudeschko, and
L Jäger, and
O Cromwell, and
H Fiebig
November 2000,
Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology,
B Fahlbusch, and
W D Müller, and
O Rudeschko, and
L Jäger, and
O Cromwell, and
H Fiebig
January 1985,
International archives of allergy and applied immunology,
B Fahlbusch, and
W D Müller, and
O Rudeschko, and
L Jäger, and
O Cromwell, and
H Fiebig
December 1993,
The Journal of allergy and clinical immunology,
B Fahlbusch, and
W D Müller, and
O Rudeschko, and
L Jäger, and
O Cromwell, and
H Fiebig
January 1988,
Allergologia et immunopathologia,
B Fahlbusch, and
W D Müller, and
O Rudeschko, and
L Jäger, and
O Cromwell, and
H Fiebig
January 1990,
International archives of allergy and applied immunology,
B Fahlbusch, and
W D Müller, and
O Rudeschko, and
L Jäger, and
O Cromwell, and
H Fiebig
June 1999,
Indoor air,
B Fahlbusch, and
W D Müller, and
O Rudeschko, and
L Jäger, and
O Cromwell, and
H Fiebig
May 1986,
Journal of immunological methods,
B Fahlbusch, and
W D Müller, and
O Rudeschko, and
L Jäger, and
O Cromwell, and
H Fiebig
January 2004,
Clinical allergy and immunology,
B Fahlbusch, and
W D Müller, and
O Rudeschko, and
L Jäger, and
O Cromwell, and
H Fiebig
January 2008,
Clinical allergy and immunology,
