Immuno-chemiluminescense detection of allergenic proteins from rubber gloves and natural rubber latex Deteksi protein allergen secara imunokimia dari sarung tangan dan lateks karet alam

Abstract

Abstrak

Lateks alam maupun produk jadi yang berasal dari karet alam diketahui mengandung protein alergen. Namun demikian identifikasi jenis protein allergen belum banyak dilaporkan. Penelitian ini bertujuan untuk mendeteksi protein alergen dari sarung tangan dan lateks karet alam menggunakan metode immuno-chemiluminescense. Protein di-ekstrak dari tiga fraksi sentrifugasi lateks (serum B, serum C dan partikel karet) serta tujuh jenis sarung tangan komersial, kemudian dipisahkan berdasarkan berat molekulnya melalui Gel elektroforesis 1-D (SDS PAGE) dan 2-D. Selanjutnya untuk deteksi protein allergen secara immuno-chemiluminescense dilaku-kan imunobloting menggunakan serum Ig_E tiga pasien yang terbukti positif alergi terhadap protein asal sarung tangan lateks, kemudian diwarnai dengan Sypro Ruby protein blot fluorescence. Hasil penelitian menunjukkan bahwa  berdasarkan hasil analisis Western blot one-DE sampel protein lateks menggunakan serum tiga orang tenaga medis yang terbukti positif alergi terhadap protein lateks, maka dapat diidentifikasi 14 jenis protein alergen pada sarung tangan lateks, empat diantaranya merupakan pita major yaitu Berat Molekul (BM) 35, 38, 46 dan 56 kDa. Protein allergen pada sarung tangan tersebut kemungkinan berasal dari bagian C-serum terutama protein  BM 46 dan 56 kDa ataupun campuran antara C-serum dan B-serum dari lateks karet alam. Hal ini dibuktikan bahwa dari sampel C-serum lateks dapat teridentifikasi 12 protein alergen,  empat diantaranya merupakan pita major yaitu BM 42, 46, 51 dan        56 kDa. Sedangkan dari sampel B-serum teridenti-fikasi tiga pita major dengan BM 14, 16 and 51 kDa. Hasil analisis Western blot 2-DE ekstrak protein sarung tangan menggunakan serum tiga orang tenaga medis yang terbukti positif alergi terhadap protein lateks, maka dapat diidentifikasi 12 - 13 spot protein alergen dengan pI at 4.0 to 7.0 dan yang paling dominan adalah dengan BM 23, 35, 38, 42, 45, 46 kDa.

Abstract

  Natural rubber latex and finished products derived from natural rubber is known to contain allergenic proteins. Nevertheless identification of allergenic protein has not been widely reported. This study aims to detect the protein allergens from the glove of hands and natural rubber latex using immuno-chemiluminescense. Proteins extracted from the latex centrifugation three fractions (serum B, serum C and rubber particles) as well as seven types of commercial gloves, then separated by molecular weight through 1-D gel electrophoresis (SDS PAGE) and 2-D. Furthermore, for the detection of allergen proteins in immuno-chemiluminescense performed immunoblotting using the serum IgE three patients who tested positive for allergy to latex gloves native protein, and then stained with fluorescence Sypro Ruby protein blot. The results showed that based on the results of Western blot analysis of one-DE latex proteins using serum samples three medical personnels who tested positive for allergy to latex proteins, we can identify 14 types of protein allergens in latex gloves, four of which are major bands that having Molecular Weight (MW) 35, 38, 46 and 56 kDa. Protein allergen on the gloves are likely to come from the C-serum protein mainly MW 46 and 56 kDa, or a mixture of C-serum and B-serum of natural rubber latex. It was proved that from C-serum samples could be identified as many as 12 protein latex allergens, four of which were major bands that MW 42, 46, 51 and 56 kDa. While the B-serum samples identified three major bands with MW 14, 16 and 51 kDa. Results of Western blot analysis of 2-DE protein extracts glove using the serum three medical personnel who tested positive for allergy to latex proteins, it could be identified 12-13 allergen protein spot with pI at 4.0 to 7.0 and most dominant is the MW 23, 35, 38, 42, 45, 46kDa.