• Catalog No.
• Service Name
• Size
• List Price (US$)
• Quantity
•  

• BS0045-1
• Cloning and sequencing hybridoma antibody heavy and light chain variable regions
• 1 hybridoma
• 1500.00

• BS0045-2
• Cloning and sequencing hybridoma antibody heavy and light chain variable regions + 100 microgram scale transient production of reconstructed scFv-Fc
• 1 package
• 4000.00

• BS0045-3
• Cloning and sequencing hybridoma antibody heavy and light chain variable regions + 100 microgram scale transient production of reconstructed whole IgG
• 1 package
• 4000.00

• BS0045-6
• Cloning and sequencing hybridoma antibody heavy and light chain variable regions + 100 ug production of reconstructed scFv-Fc + antigen-antibody binding confirmation using scFv
• 1 package
• 7000.00

• BS0045-7
• Cloning and sequencing of hybridoma antibody + 100 ug production of reconstructed whole IgG + antigen-antibody binding confirmation using whole IgGs
• 1 package
• 7000.00

Prices applied to mouse IgG antibodies. An industry record for antibody cloning and sequencing: 4 days. Prices are higher for cloning and sequencing rat, human, and primate antibodies.

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:

100000 cells per hybridoma are sufficient for cloning and sequencing the antibody variable regions (VH and VL). Welcome to challenge us with lower number of cells or non-viable cells for hybridoma rescue. (Higher fees may be charged. Please ask for a quotation.)

Turnaround:
1-2 weeks for cloning and sequencing antibody variable regions of hybridoma cell lines. An industry record: 4 days.
2-3 weeks for molecular construction, production and purification of IgG or scFv-Fc from mammalian cells, such as CHO and HEK 293.
1-2 weeks for assessment of the binding affinity and specificity of IgG or scFv.

The lowest prices are guaranteed. If you find any US company providing the same quality services with lower prices, we will return the difference to you.

So far, none of our customers has been dissatisfied with efficiency, quality or price of our antibody cloning and sequencing services.

Description:

The antibody variable regions (including both VL and VH) of hybridoma cell lines will be PCR amplified and sequenced. Antigen-antibody binding will be confirmed by transiently expressing single-chain antibodies (scFv) or whole IgGs in mammalian expression systems and testing whether the recombinant scFv or whole IgG retains the antigen binding affinity and specificity.

1. Cloning and sequencing antibody variable regions (including both VL and VH) of hybridoma cell lines:

Extract and purify total RNAs from hybridoma cells;
Synthesize cDNAs using optimized primer sets;
Amplify the DNA fragments corresponding to the VL and VH using nested PCR;
Sequence the amplified DNA fragments.
Deliverable:
A. Amplified DNA fragments
B. Sequencing results of the amplified DNA fragments
C. Summary report

2. Production of 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;
Deliverable:
Purified scFv-Fc protein (100 ug; inquire if you need more);

3. Production of 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 recombinant IgGs.
Deliverable:
Purified antibody (100 ug; inquire if you need more);

4. Antigen binding confirmation of scFv-Fc or whole IgG:

Test whether the recombinant scFv-Fc or whole IgG retains the immunological activities of their corresponding parent monoclonal antibodies.
Deliverable:
Summary report including antigen binding confirmation data.

Related Links:

• Custom Mouse Monoclonal Antibody Development Services $2800+
• Custom Recombinant Antibody Production Services
• Generation of Stable Cell Lines Expressing Antibodies
• Full Services of Custom Antibody Production and Applications
• Antibody cloning and sequencing service for $1500 only