Before cell fusion, blood samples o

Before cell fusion, blood samples of mice were
collected and its specific antibody was determined
by ELISA method using pre-coated of 96 wells ELISA
plates with recombinant OMPw. The mouse with the
best antibody titer was selected for cell fusion and
hybridoma cell generation. Briefly, Splenocytes from the
immunized mice were mixed with murine myeloma
Sp2/0 cell line at a ratio of 1:5 (1 Sp2/0 and 5 spleen cells). Cell fusion and selecting hybridoma cell was
performed in usual manner. The reactivity of hybridoma supernatants was determined using 96-wells pre-coated
ELISA plates (Nunc, Roskilde, Denmark) with 50 μL of
10 μg mL−1 of Phosphate-Buffered Saline (PBS)
dissolved rOMPw. Finally, positive hybridoma cells
were cloned 4 times by limiting dilution to select the
stable hybridomas.

Hasil (Bahasa Indonesia) 1:

Sebelum Fusi sel, sampel darah tikus yangdikumpulkan dan antibodi tertentu yang ditentukanoleh ELISA metode menggunakan pra-dilapisi 96 sumur ELISApiring dengan OMPw rekombinan. Mouse dengantiter antibodi terbaik dipilih untuk Fusi sel danhibridoma sel generasi. Secara singkat, Splenocytes daritikus diimunisasi dicampur dengan tumore myelomaSP2 0 sel baris pada rasio 1:5 (Sp2/0 dan 5 1 limpa sel). Fusi sel dan memilih hibridoma seldilakukan dengan cara biasa. Reaktivitas hibridoma supernatants ditentukan menggunakan pra-dilapisi 96-sumurELISA piring (Kidung-Kidung, Roskilde, Denmark) dengan 50 μL dari10 μg mL−1 dari Phosphate-Buffered Saline (PBS)rOMPw terlarut. Akhirnya, sel-sel hibridoma positifitu kloning 4 kali dengan membatasi pengenceran untuk memilihhybridomas stabil.