Author: Lyoo, Kwang-Soo; Yeom, Minjoo; Kim, Jungho; Kim, Donghyuk; Ha, Gunwoo; Na, Woonsung; Le, Van Phan; Song, Daesub
Title: Development of rapid immunochromatographic strip test for the detection of porcine epidemic diarrhoea virus Document date: 2017_12_2
ID: 4szmu1dh_5
Snippet: To produce monoclonal antibodies, BALB/c mice (six to eight weeks old) were intraperitoneally immunised with the purified N protein antigen emulsified in complete Freund's adjuvant. Three identical boosters were administrated with the N protein emulsified with incomplete Freund's adjuvant one-week intervals. The immunised mice were sacrificed to isolate spleen cells, which were washed in serum-free cell culture medium. Splenocytes (1×10 8 ) were.....
Document: To produce monoclonal antibodies, BALB/c mice (six to eight weeks old) were intraperitoneally immunised with the purified N protein antigen emulsified in complete Freund's adjuvant. Three identical boosters were administrated with the N protein emulsified with incomplete Freund's adjuvant one-week intervals. The immunised mice were sacrificed to isolate spleen cells, which were washed in serum-free cell culture medium. Splenocytes (1×10 8 ) were mixed with myeloma cells line Sp2/0-Ag-14 (ATCC CRL 1581) (2×10 7 ) in a ratio of 5:1 in the presence of polyethylene glycol for cell fusion. The fused cells were seeded into 96-well tissue culture plates with hypoxanthine-aminopterin-thymidine (HAT) selection medium. Culture medium was exchanged with fresh HAT selection medium eight days after fusion. On day 11, the supernatants were screened by ELISA, and then mAb anti-PED 3B12-1A6 and mAb anti-PED 1H12-1C6 hybridomas, which strongly reacted with the PEDV protein, were selected. Each hybridoma cell was intraperitoneally injected into naïve BALB/C mice to generate ascetic fluid. The ascetic fluid was then purified on protein G Sepharose 4 Fast Flow (GE Healthcare, Uppsala, Sweden) according to the manufacturer's protocol. The antibody concentrations were measured using the Bradford protein assay (Bio-Rad Laboratories, Hercules, CA, USA).
Search related documents:
Co phrase search for related documents- cell culture and fast flow: 1
- cell culture and hat selection: 1
- cell culture and hat selection medium: 1
- cell culture and hybridoma cell: 1, 2, 3, 4
- cell culture and hypoxanthine aminopterin thymidine: 1, 2, 3
- cell culture and medium selection: 1
- cell culture and monoclonal antibody: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29
- cell culture and myeloma cell: 1, 2
- cell culture and myeloma cell line: 1
- cell culture and PEDV protein: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22
- cell culture and serum free cell culture: 1, 2, 3, 4
- cell culture and serum free cell culture medium: 1, 2, 3
- cell culture and spleen cell: 1, 2, 3, 4, 5
- cell culture and tissue culture: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75
- cell culture and tissue culture plate: 1, 2
- cell fusion and culture medium: 1, 2, 3, 4, 5, 6, 7, 8, 9
- cell fusion and fuse cell: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13
- cell fusion and hat selection: 1
- cell fusion and hat selection medium: 1
Co phrase search for related documents, hyperlinks ordered by date