Abstract
Purification yolk immunoglobulin of hens vaccinated against excretory/secretory Ascaridia galli L3 larvae stageABSTRACT. The main immunoglobulin fraction of poultry is called IgY, in order to distinguish it from the mammalian IgG. This article focus on purification yolk immunoglobulin of hens vaccinated against excretory/secretory Ascaridia galli larvae to obtained purity IgY. Active vaccinations with excretory/secretory antigen were applied intra muscularly of chickens with an initial dose of 80 μg. The vaccinations were repeated three times with dose of each 60 μg with an interval of one week. The first vaccinations were excretory/secretory antigen mixed with Fruend Adjuvant Complete and subsequently mixed with Freund Adjuvant Incomplete. Antibody response in yolk was detected at weekly intervals by agar gel precipitation test (AGPT). The chicken’s eggs were collected from 49 day after vaccinations. IgY was extracted from egg yolks by means of ammonium sulphate and purified using fast performance liquid chromatography (FPLC). The purity of anti-ekscretory/secretory IgY protein was determined by Bradford method (λ = 280 nm). The result showed that antibody in yolk was begun detect with AGPT at four weeks after vaccination. IgY concentration after purification was 0,875 ± 0.251 mg/ml. This study has shown that the product released in vitro by L3 stage A. galli is capable of stimulating humoral immunity by mean of producing antibody through yolk as biologic manufacturer could be a good choice.
Highlights
Kuning telur dari ayam yang diimunisasi sudah sangat terkenal sebagai salah satu sumber antibodi
This study has shown that the product released in vitro by L3 stage A. galli is capable of stimulating humoral immunity by mean of producing antibody through yolk as biologic manufacturer could be a good choice
Http://www.mcmaster.ca/inabis98/imm unology/-szabo0509/index.html (20-1106) Yoshihara, S., Oya, T., Furuya, T., Goto, N
Summary
Rancangan Penelitian Penelitian ini menggunakan enam ekor ayam petelur jenis Hysex Brown yang dibagi atas dua kelompok. Kelompok kedua terdiri dari tiga ekor ayam yang divaksin dengan 260 g ekskretori/ sekretori L3 A. galli secara intramuskular. Apabila uji AGPT sudah menunjukkan hasil positif, maka telur ayam dikoleksi untuk dilakukan purifikasi IgY melalui Fast Performan Liquid Chromatography (FPLC). Konsentrasi protein antigen ekskretori/ sekretori L3 A. galli dosis 260 g ditentukan berdasarkan uji Bradford seperti dijelaskan oleh Darmawi et al (2009) digunakan pada penelitian ini. Reaksi dibaca setelah 18 – 48 jam untuk melihat adanya garis presipitasi yang menunjukkan antara antibodi di dalam kuning telur dan antigen ekskretori/ sekretori L3 A. galli tersebut terjadi reaksi homolog (Lanyi dan Bergan, 2003). Kuning telur dari ayam yang telah divaksinasi dengan antigen asal L3 A. galli diencerkan dengan 9 bagian aquadest pH 5,0 – 5,2 dan diinkubasikan selama 6 jam pada suhu 4oC. Absorban sampel ditentukan dengan pembacaan panjang gelombang 280 nm (Paryati, 2006)
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