Produksi antibodi monoklonal terhadap M-Like protein Streptococcus equi subsp. zooepidemicus isolat monyet ekor panjang (Macaca fascicularis

PURWANTORO, Aris, Dr. Widya Asmara

2005 | Tesis | S2 Bioteknologi

Infeksi Streptococcus equi subsp. zooepidemicus yang menyerang babi, monyet ekor panjang, dan manusia memerlukan penanganan intensif. Penelitian ini bertujuan memproduksi antibodi monoklonal terhadap Mlike protein (MLP) bakteri tersebut, sehingga deteksinya dapat cepat, akurat dan massal. Identifikasi MLP dilakukan terhadap tiga isolat Streptococcus equi subsp. zooepidemicus (6.70, 6.72, dan 6.73) yang diisolasi dari monyet ekor panjang di Pulau Bali. Protein MLP 58 kDa dari isolat 6.70 dipakai sebagai bahan pembuatan antibodi monoklonal. Imunisasi mencit Balb/c dengan protein MLP 58 kDa dilakukan sekali menggunakan Freund’s complete adjuvant dan dilanjutkan Freund’s incomplete adjuvant sebanyak tiga kali. Fusi sel dilakukan antara sel limfosit mencit imun dan sel mieloma (P3-NS1-Ag4-1). Hasil penelitian diperoleh empat klon hibridoma yang menghasilkan antibodi monoklonal terhadap protein MLP 58 kDa. Keempat antibodi monoklonal menunjukkan sensitivitas yang tinggi dalam mendeteksi infeksi Streptococcus equi subsp. zooepidemicus pada monyet ekor panjang.

The infection of Streptococcus equi subsp. zooepidemicus on pigs, long-tail macaques, and human needs an intensive and better handling method. This research was carried out to produce monoclonal antibody against M-like protein (MLP) of the bacterium, so the detection of the infection will be more accurate, faster, and able to assay mass samples. Three isolates of Streptococcus equi subsp. zooepidemicus (6.70, 6.72, and 6.73) which isolated from long-tail macaques in Bali Island were identified for their MLP. The MLP 58 kDa of 6.70 isolate was used to produce monoclonal antibody. Balb/c mice was immunized once with the MLP 58 kDa and Freund’s complete adjuvant and continued three times with mixture of the MLP 58 kDa and Freund’s incomplete adjuvant. The immune lymphocyte cells were fused with myeloma cells (P3-NS1-Ag4-1). The result shows that four clones of hybridomas could produced monoclonal antibody against MLP 58 kDa. The monoclonal antibody shows high sensitivity to detect Streptococcus equi subsp. zooepidemicus infection on long-tail macaques.

Kata Kunci : Streptococcus equi subsp. zooepidemicus, Macaca fascicularis, monyet ekor panjang, M-like protein, antibodi monoklonal, long-tail macaque, monoclonal antibody