Mouse/human chimeric IgG1 and IgG4 antibodies directed to the house dust mite allergen Der p 2: use in quantification of allergen specific IgG.
OBJECTIVE
Chimeric mouse/human monoclonal IgG1 and IgG4 antibodies were developed against the house dust mite allergen Der p 2. These chimeric IgG antibodies, hIgG1-Dp2 A and hIgG4-Dp2 A, have the same binding characteristics as the previously reported chimeric hIgE-Dp2 A and are composed of the heavy chain variable domains and light chains of the original murine monoclonal antibody 2B12, whereas the heavy chain constant domains have been replaced by the human IgG1 or IgG4 heavy chain. The expression level of hIgG1-Dp2 A and hIgG4-Dp2 A was 1 and 3.5 microg/mL, respectively.


RESULTS
Since all IgG in these culture supernatants is allergen-specific, they are useful reference reagents and enable the calculation of the amount of allergen specific IgG1 and IgG4 antibodies in absolute IgG amounts. The results obtained with two panels of sera from patients in immunotherapeutic treatment were evaluated and compared in Der p 2 IgE, IgG1 and IgG4 RAST and with reversed IgG4 RAST using labelled purified Der p 2. Close agreement between the results for the two IgG4 assays was found.


CONCLUSIONS
With these chimeric reference reagents the quantities of isotype specific antiallergen antibodies can be calculated and compared.
Schuurman
J
J
Central Laboratory of the Red Cross Blood Transfusion Service and Laboratory of Experimental and Clinical Immunology, University of Amsterdam, The Netherlands.
Perdok
G J
GJ
Mueller
G A
GA
Benjamin
D C
DC
Yong Tan
K
K
Chapman
M D
MD
Aalberse
R C
RC
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D039741:Antigens, Dermatophagoides; D002460:Cell Line; D004391:Dust; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D006825:Hybridomas; D006967:Hypersensitivity / Q000628:therapy; D007073:Immunoglobulin E / Q000097:blood; D007074:Immunoglobulin G / Q000097:blood* / Q000276:immunology / Q000378:metabolism; D007167:Immunotherapy; D051379:Mice; D008925:Mites; D011852:Radioallergosorbent Test; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D039741:Antigens, Dermatophagoides; D004391:Dust; D006023:Glycoproteins; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D007073:Immunoglobulin E