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Clonacell hy kit

Manufactured by STEMCELL
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The ClonaCell™-HY Kit is a laboratory product designed for the isolation and clonal expansion of hybridoma cells. It provides the necessary components to support the growth and selection of hybridoma cells, which are used for the production of monoclonal antibodies.

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Lab products found in correlation

Quil a, by Merck Group (1 mentions) Protein g affinity chromatography, by GE Healthcare (1 mentions)

4 protocols using clonacell hy kit

1

Hybridoma Cells Production for CBPP Antibodies

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The BALB/c mice (n = 4, 4 weeks old) were intraperitoneally (IP) immunized with 200 µl of crude CBPP T1/44 antigen emulsion at a concentration of 0.3 mg/ml together with the Freund's complete adjuvant mixed at a 1 : 1 ratio for antibodies production against CBPP antigen, as described by [13 (link), 14 (link)]. Booster injections were administered every two weeks. The first three booster injections were composed of the CBPP antigen and incomplete adjuvant mixed at the same ratio as the first injection. The last booster injection (fourth) was 200 µl crude CBPP antigen in saline without adjuvant. Four days after the last boost, the mice were sacrificed, and the spleen cells were prepared (as described by [15 (link)]; StemCell Technologies, ClonaCell™-HY Kit, Canada) for fusion with the tumor Ag8 myeloma cells (FAO/IAEA Laboratories, Austria) (prepared as described by [16 (link)] and StemCell Technologies, ClonaCell™-HY Kit, Canada) for the production of hybridoma cells. The hybridoma fusion, selection, and harvesting were carried out step by step according to the ClonaCell™-HY Kit (StemCell Technologies, Canada) [15 (link)].
Ramathudi-Dunbar L., Awosanya E., Charles S.B., Chitsungo E., Moustapha Boukary C.R., Nwankpa N., Gelaw H., Tessema Y., Melesse A. G., Rayson Sanga R., Bright A, & Baziki J.D. (2024). Development and Evaluation of Monoclonal Antibodies against CBPP Antigen with the End Goal of Developing an ELISA Kit. Veterinary Medicine International, 2024, 6901355.
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2

Generation of TCRm Monoclonal Antibodies

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Female BALB/c mice were immunized with a solution of 50 μg of purified peptide/HLA-A*02:01 complexes and Quil-A adjuvant (Sigma) at 15 day intervals as described (23 (link), 24 (link)). One week after the third immunization, serum samples were evaluated for polyclonal TCRm mAb responses. Mice that had responses showing a signal to noise ratio of > 2 fold in absorbance reading and a titer of > 1/600 were selected for hybridoma generation. Hybridomas were produced by fusing splenocytes with the P3X63.Ag8.653 myeloma cell line using the clonacell-HY Kit (Stem Cell Technologies). After two weeks in semi-solid medium, single clones were picked, transferred to 96-well tissue culture plates and grown for 3–4 days. Hybridoma supernatants were screened for specific mAb production by ELISA and cell-based flow cytometric assays. Hybridoma lines were cultured in Hybridoma-SFM (Serum Free Media) (Gibco) and the RL14C (anti SLF9/HLA-A2), RL15A (anti SVG9/HLA-A2) and RL26A (anti YTM9/HLA-A2) TCRm mAbs were purified from cell supernatants by affinity chromatography with protein A Sepharose (Amersham-GE, Boston, MA).
Kaabinejadian S., McMurtrey C.P., Kim S., Jain R., Bardet W., Schafer F.B., Davenport J.L., Martin A.D., Diamond M.S., Weidanz J.A., Hansen T.H, & Hildebrand W.H. (2016). Immunodominant West Nile Virus T Cell Epitopes Are Fewer in Number and Fashionably Late. Journal of immunology (Baltimore, Md. : 1950), 196(10), 4263-4273.
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3

Generation of Anti-Citrullinated Peptide Antibodies

Cited 1 time
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BALB/c and C57BL/6 mice were immunized with chemically modified (*) citrulline peptides (PIECit*TYLK-NH2 and YAGCit*LLTK-NH2), coupled to either keyhole limpet hemocyanin (KLH) or ovalbumin. Different citrulline-containing peptides were created in the host laboratory via solid phase peptide synthesis, as described previously (20 (link)). Hybridoma cell lines were created using the ClonaCell®-HY kit (Stemcell Technologies) according to the manufacturer’s instructions. SP2/0 mouse myeloma cells were fused with purified splenocytes of immunized mice. Stable hybridomas were screened and selected on the basis of antibody reactivity to different synthetic citrullinated and control peptides that were coated on the plates at 100 ng/ml with the use of indirect ELISAs (20 (link)). Further validation was performed by Western Blot analysis and the use of synthetic citrullinated and non-citrullinated CXCL8 isoforms (21 (link)). Cell culture supernatants were filtered and antibodies were purified on Protein G affinity chromatography (GE Healthcare).
Grillet B., Yu K., Ugarte-Berzal E., Janssens R., Pereira R.V., Boon L., Martens E., Berghmans N., Ronsse I., Van Aelst I., Fiten P., Conings R., Vandooren J., Verschueren P., Van Damme J., Proost P, & Opdenakker G. (2021). Proteoform Analysis of Matrix Metalloproteinase-9/Gelatinase B and Discovery of Its Citrullination in Rheumatoid Arthritis Synovial Fluids. Frontiers in Immunology, 12, 763832.
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4

Generating Anti-SLAMF7 Monoclonal Antibodies

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Example 1

In this study, a panel of monoclonal antibodies (mAbs) was raised against SLAMF7 (CD319), a cell surface marker that is associated to MM. To generate these mAbs, 4 female BALB/c mice were immunized with recombinant human SLAMF7 (CD319) purchased from Sino Biological Inc. (Catalogue No. 11691-H08H) (FIG. 1). The DNA sequence that encodes the recombinant human SLAM7 (CD319), consists of an extracellular domain (Met1 to Met226) fused with a polyhistidine tag at the C-terminus, that was expressed via human host cells.

Mice immunization started on 6 Feb. 2014, and this was followed by an immunisation regime consisting of 5 weekly immunisations and 1 booster immunisation. Upon completion of the immunisation regime, plasma B cells were harvested and fused with SP2/0 mouse myeloma lines on 13 Mar. 2014 (FIG. 1). Hybridoma fusion was performed using STEMCELL Technologies ClonaCell™-HY kit and generated a panel of hybridomas that produces mAbs present in the culture supernatant. The mAbs were screened by flow cytometry for binding with AML, MM, other cancer and normal cell lines, that subsequently identified a novel anti-SLAMF7 antibody, SLAM01.

US11124579B2. Anti oligosaccharide antibody (2021-09-21). AGENCY FOR SCIENCE, TECHNOLOGY AND RESEARCH [SG], NATIONAL UNIVERSITY OF SINGAPORE [SG]. Inventors: Chong Kok Teo [SG], Andre Boon Hwa Choo [SG], Wee Joo Chng [SG].
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