Monoclonal antibodies against the candidate lupin allergens α-conglutin and β-conglutin

被引:26
作者
Dooper, Maaike M. B. W.
Holden, Lise
Faeste, Christiane K.
Thompson, Keith M.
Egaas, Eliann
机构
[1] Natl Vet Inst, NO-0033 Oslo, Norway
[2] Univ Oslo, Inst Immunol, Oslo, Norway
[3] Rikshosp Univ Hosp, Oslo, Norway
关键词
conglutins; enzyme-linked immunosorbent assay; globulins; lupin; monoclonal antibodies;
D O I
10.1159/000098224
中图分类号
R392 [医学免疫学];
学科分类号
100102 ;
摘要
Background: The ingestion of dietary products containing sweet lupin (such as Lupinus albus or Lupinus angustifolius) has been reported to cause IgE-mediated allergic reactions. Recent studies have indicated lupin globulins as important IgE binding proteins. The aim of the present study was to generate and characterize monoclonal antibodies (mAbs) against lupin seed proteins. Methods: Mice were immunized with a protein isolate from L. albus and mAbs were obtained by hybridoma techniques. Albumins and globulins were extracted, and the globulin fraction was separated further into conglutins by anion exchange chromatography. Specificities, binding patterns and applications of the mAbs were investigated by immunochemical methods. Results: Five mAbs were produced: Lu11 (an IgG2b antibody), Lu8, Lu18, Lu34 and Lu35 (all IgM antibodies). The mAbs reacted strongly with protein isolates from both L. albus and L. angustifolius. All mAbs are directed towards the lupin globulin fraction; Lu11 and Lu18 recognize alpha-conglutin, while Lu8, Lu34 and Lu35 recognize beta-conglutin. In addition, Lu11 inhibited the binding of IgE from patients with positive skin prick tests to lupin proteins in a competitive ELISA by approximately 30%. Furthermore, preliminary results show that Lu11 can be used to develop a sensitive method for the detection of beta-conglutin in foods. Conclusions: Lupin globulins are immunogenic and beta-conglutin is a potential allergen. This is the first study describing mAbs against the candidate lupin allergens, emphasizing the importance of additional studies on conglutins in lupin allergy.
引用
收藏
页码:49 / 58
页数:10
相关论文
共 37 条
[1]   Detection of recombinant Alt a1 in a two-site, IgM based, sandwich ELISA opens up possibilities of developing alternative assays for the allergen [J].
Abebe, Makonnen ;
Kumar, Veena ;
Rajan, Soundaram ;
Thaker, Ajay ;
Sevinc, Serdal ;
Vijay, Hari M. .
JOURNAL OF IMMUNOLOGICAL METHODS, 2006, 312 (1-2) :111-117
[2]   Effects of extrusion, boiling, autoclaving, and microwave heating on lupine allergenicity [J].
Alvarez-Alvarez, J ;
Guillamón, E ;
Crespo, JF ;
Cuadrado, C ;
Burbano, C ;
Rodríguez, J ;
Fernández, C ;
Muzquiz, M .
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 2005, 53 (04) :1294-1298
[3]  
BLAGROVE R J, 1975, Australian Journal of Plant Physiology, V2, P13
[4]  
Breiteneder H, 2004, J ALLERGY CLIN IMMUN, V113, P821, DOI [10.1016/j.jaci.2004.01.779, 10.1016/j.jaci.2004.01.079]
[5]  
DEPENNA EW, 1987, J FOOD SCI, V52, P1434
[6]   HEAT-INDUCED SYNTHESIS AND TUNICAMYCIN-SENSITIVE SECRETION OF THE PUTATIVE STORAGE GLYCOPROTEIN CONGLUTIN-GAMMA FROM MATURE LUPIN SEEDS [J].
DURANTI, M ;
SCARAFONI, A ;
GIUS, C ;
NEGRI, A ;
FAORO, F .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1994, 222 (02) :387-393
[7]   THE SEED GLOBULINS OF LUPINUS-ALBUS [J].
DURANTI, M ;
RESTANI, P ;
PONIATOWSKA, M ;
CERLETTI, P .
PHYTOCHEMISTRY, 1981, 20 (09) :2071-2075
[8]   Calcium- and magnesium-dependent aggregation of legume seed storage proteins [J].
Ferreira, RB ;
Franco, E ;
Teixeira, AR .
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 1999, 47 (08) :3009-3015
[9]   Utilization of an improved methodology to isolate Lupinus albus conglutins in the study of their sedimentation coefficients [J].
Franco, E ;
Ferreira, RB ;
Teixeira, AR .
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 1997, 45 (10) :3908-3913
[10]   A case of peanut cross-allergy to lupine flour in a hot dog bread [J].
Fæste, CK ;
Lovik, M ;
Wiker, HG ;
Egaas, E .
INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY, 2004, 135 (01) :36-39