scholarly journals Production of monoclonal antibodies against conserved components of infectious bronchitis virus

2001 ◽  
Vol 53 (5) ◽  
pp. 523-530 ◽  
Author(s):  
C.M. Souza ◽  
F.R.T. Rocha ◽  
N.R.S. Martins ◽  
J.S. Resende ◽  
M.A. Jorge ◽  
...  
Keyword(s):  
Differential Diagnosis ◽  
Monoclonal Antibodies ◽  
Cell Line ◽  
Western Blotting ◽  
Infectious Bronchitis Virus ◽  
Myeloma Cell ◽  
Spleen Cells ◽  
Myeloma Cell Line ◽  
Infectious Bronchitis ◽  
Routine Diagnosis

Murine hybridomas producing IgG1 monoclonal antibodies (Mabs) against N and S2 proteins (53KDa and 82KDa, respectively) from avian infection bronchitis virus (IBV) strain M41 were generated by the fusion of a myeloma cell line (Sp2/0-Ag14) with spleen cells from Balb/c mice previously immunized with whole virus IBV M41. Post-fusion screening criterion was by ELISA and 36 positive hybrids were generated after fusions. Two hybrids specific to N (N3F10) and S2 (S12B2) proteins from M41 (serotype Massachusetts) were selected by western blotting. These Mabs recognized the Ark-99 (serotype Arkansas) and A5968 (serotype Connecticut) IBV strains in addition to M41. By ELISA, the Mab against the S2 (S12B2) recognized all reference and Brazilian strains (M41, SE-17, H52, 297, 283, PM-1, PM-2, PM-3, 351, 29-78 E 327) studied, while the Mab against N recognized only six (M41, SE-17, H52, 283, 327 e 297) strains. The Mab against S2 may become a useful tool for IBV detection on the routine diagnosis of infectious bronchitis, especially for helping the differential diagnosis of clinically and pathologically confusing diseases, while the Mab against N (N3F10) recognized a probably less conserved region among the strains and may be interesting to comparing IBV isolates.

scholarly journals Preparation of Hybridomas Producing Monoclonal Antibodies against Aflatoxin B1 as a Tool to Control Hepatocellular Carcinoma

2019 ◽  
Vol 13 ◽  
pp. 1-12
Author(s):  
Manal M.E. Ahmed ◽  
Rafik Soliman ◽  
Jakeen Eljakee ◽  
Ahmed El-Sanousi ◽  
Haitham Amer ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Aflatoxin B1 ◽  
Quantitative Measurement ◽  
Myeloma Cell ◽  
Selective Medium ◽  
Immunization Schedule ◽  
Spleen Cells ◽  
Myeloma Cell Line ◽  
High Affinity ◽  
Multiple Uses

Hybridomas that secreted antibodies against aflatoxin B1 for multiple uses were prepared using a unique immunization schedule. Aflatoxin B1-BSA conjugate was used for immunization of Balb/c mice. Spleen cells were harvested from the hyper immunized mice to be fused with myeloma cell line (P3NS1) using polyethylene glycol 3000, 50% concentration as a fusogenic agent. The produced hybridomas were selected using HAT selective medium that was replaced by complete HT medium. From the 10thday after fusion, wells that contain colonies of hybridomas covering 30% or greater of the wells surface were screened for production of monoclonal antibodies against aflatoxin B1 using ELISA. 21 hybridomas were found to be reactive to aflatoxin B1. All were found to belong to IgG2aisotype except one was found to belong to IgM isotype. The prepared monoclonal antibodies and their application to immunoassays represents a useful and rapid quantitative measurement with high affinity and low detection limits in order to purify environmentally occurring levels of this carcinogen specially in areas at high risk for liver cancer.

scholarly journals Production and Characterization of Monoclonal Antibodies to Psittacine Beak and Feather Disease Virus

1992 ◽  
Vol 4 (1) ◽  
pp. 13-18 ◽  
Author(s):  
Branson W. Ritchie ◽  
Frank D. Niagro ◽  
Kenneth S. Latimer ◽  
W. L. Steffens ◽  
Denise Pesti ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Myeloma Cell ◽  
Enzyme Linked Immunosorbent Assay ◽  
Spleen Cells ◽  
Myeloma Cell Line ◽  
Elisa System ◽  
Mouse Myeloma Cell Line ◽  
Feather Disease Virus ◽  
Mouse Myeloma

Monoclonal antibodies specific for the virus that causes psittacine beak and feather disease (PBFD) were produced by fusing spleen cells from mice immunized with purified concentrated PBFD virus with mouse myeloma cell line Sp2/0. The resulting hybridomas were tested for reactivity against whole purified virus by an enzyme-linked immunosorbent assay (ELISA) system. Four clones, designated 15H8, 8E3, 11G12, and 2C3, were subcloned by limiting dilution. Isotyping indicated that clone 15H8 was secreting IgG, whereas the remaining clones secreted IgM. The secreted immunoglobulins were characterized by reactivity against purified PBFD virus using immunoblotting procedures, by immunohistochemical staining of virus-induced lesions in infected tissues, and by inhibition of PBFD virus agglutination of cockatoo erythrocytes. Antibodies secreted by clones 15H8 and 8E3 had the strongest activity against purified whole virus. Only immunoglobulin secreted by the clone 15H8 could be used to detect viral antigen in infected tissues. None of the monoclonal antibodies had hemagglutination-inhibition activity.

scholarly journals The production of monoclonal antibodies to rubella haemagglutinin and their use in antibody-capture assays for rubella-specific IgM

1982 ◽  
Vol 88 (2) ◽  
pp. 335-350 ◽  
Author(s):  
R. S. Tedder ◽  
J. L. Yao ◽  
M. J. Anderson
Keyword(s):  
Monoclonal Antibodies ◽  
Single Cell ◽  
Cell Line ◽  
Ascitic Fluid ◽  
Solid Phase ◽  
Myeloma Cell ◽  
Myeloma Cell Line ◽  
Cell Clones ◽  
Antibody Capture ◽  
Limiting Dilution

SummaryMice were immunized by three intraperitoneal and one intravenous injection of rubella haemagglutinin. Splenocytes from these mice were fused with the cells of a syngeneic myeloma cell line, and following culture for various periods of time, single-cell clones were derived by the technique of limiting dilution.A total of 139 clones were derived from 13 parent hybrid cultures. To date, four of these cloned cultures have been propagated as ascitic tumours in mice. The preparation of IgG from ascitic fluid and labelling of this antibody with 125I is described. Results indicate that the use of labelled monoclonal antibodies as indicator reagents in solid-phase IgM antibody capture assays for the detection of rubella-specific IgM results in enhanced performance of these tests.

scholarly journals Production and characterization of a monoclonal antibody to biotin

1986 ◽  
Vol 237 (2) ◽  
pp. 477-482 ◽  
Author(s):  
K Dakshinamurti ◽  
R P Bhullar ◽  
A Scoot ◽  
E S Rector ◽  
G Delespesse ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Myeloma Cell ◽  
Enzyme Linked Immunosorbent Assay ◽  
Spleen Cells ◽  
Myeloma Cell Line ◽  
Biotin Deficiency ◽  
Keyhole Limpet Haemocyanin ◽  
Limiting Dilution

Monoclonal antibodies to biotin have been prepared by using biotin linked to keyhole limpet haemocyanin (KLH) as the antigen. Spleen cells obtained from mice immunized with biotin-KLH were fused with the myeloma cell line NS-1. The resulting hybridomas were screened for the production of antibodies to biotin using an enzyme-linked immunosorbent assay. Clones producing antibodies to biotin were isolated by limiting dilution methods. Four cell lines, each derived originally from a different fusion, were chosen for the production of monoclonal antibodies. The monoclonal antibodies obtained have been characterized with respect to their ability to interact with biotin, biotin-bovine serum albumin, biotin-KLH and biocytin as well as to inhibit biotin-dependent enzymes. They have been used to produce cellular biotin deficiency in vitro for studies of biotin function.

scholarly journals Isolation of monoclonal antibodies recognizing rat bone-associated molecules in vitro and in vivo.

1992 ◽  
Vol 40 (9) ◽  
pp. 1339-1352 ◽  
Author(s):  
K Turksen ◽  
U Bhargava ◽  
H K Moe ◽  
J E Aubin
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Surface ◽  
Myeloma Cell ◽  
Spleen Cells ◽  
Myeloma Cell Line ◽  
Osteogenic Cells ◽  
Fetal Rat ◽  
Osteoblast Lineage

Knowledge of the number and kinds of differentiation steps that characterize cells of the osteoblast lineage is inadequate. To further analyze osteoblast differentiation, we generated a series of monoclonal antibodies (MAb) to osteogenic cells. Spleen cells from mice immunized with whole-cell populations enriched for expression of osteoblast-associated properties or bone formation in vitro were fused with the SP2/0 myeloma cell line. Supernatants from growing hybridomas were screened by indirect immunofluorescence on frozen sections of a portion of 21-day fetal rat heads that included the calvaria bone, periosteum, muscle, fibrous connective tissue, and skin. Six MAb were selected with bone-associated staining and limited ability to label other tissues. Either cell surface or cytoplasmic molecules were recognized by five of the MAb; one recognized a molecule detectable both in the cytoplasm, on the cell surface, and in the extracellular matrix. Of the antibodies selected, one identified both preosteoblasts and osteoblasts and has been found to be against alkaline phosphatase. The others recognized the mature osteoblasts, osteocytes, and chondrocytic cells. The pattern and distribution of the labeling in vivo extended to primary cells and cell lines in vivo. These results support earlier observations on molecules differentially expressed by cells at different stages of the osteoblast lineage and extend the available cell surface and cytoplasmic epitopes identifiable as marker molecules.

Use of monoclonal antibodies for radioimmunoassay of water buffalo milk progesterone

1987 ◽  
Vol 54 (4) ◽  
pp. 471-477 ◽  
Author(s):  
Rosanna Capparelli ◽  
Domenico Iannelli ◽  
Aldo Bordi
Keyword(s):  
Monoclonal Antibodies ◽  
Water Buffalo ◽  
Myeloma Cell ◽  
Buffalo Milk ◽  
Spleen Cells ◽  
Myeloma Cell Line ◽  
Mouse Myeloma Cell ◽  
Mouse Myeloma Cell Line ◽  
The Mean ◽  
Mouse Myeloma

SummaryIn order to standardize a radioimmunoassay of milk progesterone as a routine method for confirmation of oestrus and diagnosis of pregnancy in water buffalo, monoclonal antibodies against progesterone were produced. Hybridomas were prepared by fusing spleen cells from a Balb/c mouse immunized with progesterone 11α-hemisuccinate–bovine serum albumin conjugate with the mouse myeloma cell line NS-1. Thirty wells out of 94 secreted anti-progesterone antibodies. Of the ten independent hybridomas derived, one (AF65) was suitable for the quantification of milk progesterone by radioimmunoassay. The tracer used in the assay was progesterone-11α-hemisuccinate ([2-125I]iodohistamine). The sensitivity of the assay was 50 pg/tube. The mean progesterone concentration at oestrus was 0·8±0·2 ng/ml increasing to 8·5±0·8 ng/ml 24 d later in pregnant animals.

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