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Generation of Stable Cell Lines Expressing Antibodies
Stable Cell Line Development for Recombinant Antibody Expression and Purification
Note: Price/availability/specifications subject to change without notice. If not specified, all products and services are for in vitro research use only. Not for use in or on humans or animals.
Order offline: Phone: 1-617-401-8149, Fax: 1-617-606-5019, Email: message@sydlabs.com, or leave a message with a formal purchase order (PO) or credit card.
Features:
Turnaround:
1-2 weeks for cloning and sequencing antibody variable regions of hybridoma cell lines.
2 weeks for synthesizing antibody variable regions.
2-3 weeks for molecular construction of IgG or scFv-Fc.
1 week for assessment of the binding affinity and specificity of IgG or scFv.
2-3 weeks for generation of CHO stable cell lines expressing scFv-Fc or whole IgGs.
Experienced scientists working on antibody services for decades.
Description:
Cloned or synthesized antibody variable regions will be cloned into proprietary vectors for construction of scFv-Fc (human or mouse Fc) or whole IgGs (human constant domain or mouse constant domain), which have a heavy chain and a light chain. CHO stable cell lines expressing scFv-Fc or whole IgGs will be generated.
If requested, the antibody variable regions (including both VL and VH) of hybridoma cell lines will be cloned and sequenced. Antigen-antibody binding affinity will be confirmed by transiently expressing single-chain antibodies (scFv) or whole IgGs in mammalian expression systems and testing whether the recombinant scFv or antibodies retain the immunological activities of their corresponding parent monoclonal antibodies.
1. Antigen-antibody binding confirmation using single-chain antibodies (scFv):
Connect VL with VH using a peptide linker;
Construct scFv-Fc (human IgG1 backbone);
Transiently express recombinant scFv-Fc in mammalian expression systems;
Purify recombinant scFv-Fc;
Test whether the recombinant scFv-Fc retains the immunological activities of its corresponding parent monoclonal antibody.
Deliverable: purified scFv-Fc protein (100 ug; inquire if you need more).
2. Antigen-antibody binding confirmation using whole IgGs (human or mouse constant region IgG1):
Fuse VL and VH into human or mouse IgG constant region backbone in mammalian expression vectors;
Transiently express the recombinant antibody in mammalian expression systems;
Purify IgG and test whether the recombinant antibody retains the immunological activities of its corresponding parent monoclonal antibody.
Deliverable: purified antibody (100 ug; inquire if you need more).
3. Stable cell line development using mammalian cells:
Transfection into mammalian cells and screening for cell lines permanently expressing specific genes through their incorporation into the cellular genome. The standard stable cell line generation service by default uses monolayer CHO cells and expression vector with the zeocin selectable marker. Clones will be screened using ELISA, Western, or cell-based assay of your interest. Our faster, more predictive screening techniques may significantly increase speed of stable cell line development while improving quality and uniformity.
Deliverable:
a. 10 individual clones.
b. Stable cell line development report.
Related Links:
- Guaranteed Mouse Monoclonal Antibody Development Services
- Custom Recombinant Antibody Production Services
- Full Services of Custom Antibody Production and Applications
- Standard Services of Stable Cell Line Development
- Antibody Sequencing, Hybridoma Sequencing, Antibody Cloning
