Development of an interleukin (IL)-33 sandwich ELISA kit specific for mature IL-33.
Interleukin (IL)-33 is an inflammatory cytokine and belongs to the IL-1 family of cytokines. There are eleven members of the IL-1 family of cytokines and all have important roles in host defense against infections. Their levels are increased during infection and in various auto-inflammatory diseases. IL-33 is also associated with autoimmune diseases such as asthma, atopic dermatitis, rheumatoid arthritis, and atherosclerosis. IL-33 receptors consist of IL-1R4 and IL-1R3 to induce both Th1 and Th2 type immune response. Here we present the development of monoclonal antibodies (mAbs) against human mature IL-33. Recombinant human mature IL-33 protein was expressed in E. coli and purified by multi-step affinity chromatography. The human IL-33 activity was examined in HMC-1 and Raw 264.7 cells. Mice were immunized with the biologically active mature IL-33 to generate mAb against IL-33. The anti-IL-33 mAb (clone/4) was used as a capture antibody for a sandwich enzyme-linked immunosorbent assay (ELISA). This assay detects mature IL-33 with a high sensitivity (80 pg/mL) but does not recognize the biologically inactive precursor IL-33. This article describes the methods for a newly developed IL-33 ELISA kit that is specific for mature IL-33 and may be used to analyze bioactive mature IL-33 in various immunological diseases.
a Laboratory of Cytokine Immunology, Department of Biomedical Science and Technology , Konkuk University , Seoul , Korea.
Eunsom
E
Kim
a Laboratory of Cytokine Immunology, Department of Biomedical Science and Technology , Konkuk University , Seoul , Korea.|b College of Veterinary Medicine , Konkuk University , Seoul , Korea.
Areum
A
Kwak
a Laboratory of Cytokine Immunology, Department of Biomedical Science and Technology , Konkuk University , Seoul , Korea.|b College of Veterinary Medicine , Konkuk University , Seoul , Korea.
Hyunjhung
H
Jhun
a Laboratory of Cytokine Immunology, Department of Biomedical Science and Technology , Konkuk University , Seoul , Korea.
Siyoung
S
Lee
a Laboratory of Cytokine Immunology, Department of Biomedical Science and Technology , Konkuk University , Seoul , Korea.|b College of Veterinary Medicine , Konkuk University , Seoul , Korea.
Seunghyun
S
Jo
a Laboratory of Cytokine Immunology, Department of Biomedical Science and Technology , Konkuk University , Seoul , Korea.
Jongho
J
Lee
c Department of Biotechnology , College of Biomedical and Health Science, Konkuk University , Seoul , Korea.
Tae-Bong
TB
Kang
d Department of Immunology , College of Medicine, Konkuk University , Seoul , Korea.
Erk
E
Her
e Department of Bioequivalence Division for Drug Evaluation , Ministry of Food and Drug Safety, Konkuk University , Seoul , Korea.
Suyoung
S
Bae
f Department of Medicine, Pusan Paik Hospital, College of Medicine , Inje University , Busan , Korea.
Youngmin
Y
Lee
a Laboratory of Cytokine Immunology, Department of Biomedical Science and Technology , Konkuk University , Seoul , Korea.|b College of Veterinary Medicine , Konkuk University , Seoul , Korea.
Soohyun
S
Kim
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000937:Antigen-Antibody Reactions; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D000067596:Interleukin-33 / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D051379:Mice; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology
D000911:Antibodies, Monoclonal; C506536:IL33 protein, human; D000067596:Interleukin-33; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha
IL-33 ELISA kit; TNFα; affinity purification; cytokine; recombinant il-33