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Technical notes View data: high-throughput cloning for antibody development
antibody purification Antibody purification kits

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Technical notes View data: high-throughput cloning for antibody development
antibody purification Antibody purification kits

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Antibody therapeutics

The use of monoclonal antibodies (mAbs) for cancer therapy has achieved considerable success in recent years due to their high specificity, activity, and favorable pharmacokinetics, as well as the availability of strategies to successfully engineer antibodies into humanized forms. Antibodies are capable of recruiting the immune system to attack cancer cells through complement-dependent cytotoxicity or antibody-dependent cellular cytotoxicity (Liu and Mardis 2017). Mechanisms of direct tumor cell killing by antibody-mediated apoptosis include neutralization of cell-surface-bound enzymes and induction of receptor agonist or antagonist activity.

In another approach, antibodies deliver drugs targeting cancer cells via antibody-drug conjugates (ADCs). ADCs direct cytotoxic compounds to tumor cells after selective binding to cancer cell surface antigens, internalization, and intracellular drug release (Wold et al. 2016). Moreover, ADCs are powerful new treatment options for lymphomas and solid tumors, and immunomodulatory antibodies have recently achieved remarkable clinical success. New advances in protein engineering technology have also generated multiple bispecific antibody (BsAb) formats capable of targeting numerous antigens as a single agent and directly targeting immune cells to tumors, thus reducing drug resistance and severe adverse side effects (Thakur et al. 2018).

Highlighted products

In-Fusion Cloning technology enhances antibody discovery and engineering workflows by speeding up the generation of antibody expression constructs. In-Fusion Cloning is fast (15 min), highly efficient (>95% cloning efficiency), sequence independent (any PCR insert can be cloned into any vector at any locus), seamless (no extra bp or amino acids added to target antibody), directional, and high-throughput (HTP) ready. Several publications have utilized the In-Fusion Cloning technology for HTP antibody fragment cloning due to its high cloning efficiency and accuracy (Spidel et al. 2016; Chen et al. 2014; Meng et al. 2015).

After expression construct generation and downstream expression of the target antibody, our next-gen Capturem Protein A and Capturem Protein G technologies can be utilized for rapid (5–15 min) and easy resin-free purification of high-quality and concentrated monoclonal/polyclonal antibodies. Our revolutionary Capturem Protein A and G technologies consist of spin columns or plates containing high-capacity membranes functionalized with Protein A or G (Figure 1), allowing antibody purification directly from complex matrices such as hybridoma supernatants or serum—within minutes. Capturem Protein A and Capturem Protein G technologies are available in miniprep, maxiprep, and 96-well/24-well-plate formats, which are ideal for rapid HTP screening of hybridoma clones.

Capturem Protein A/G technology

Figure 1. Capturem Protein A and Protein G technology. Capturem Protein A and G mini/maxi spin columns or 96-well/24-well plates contain a high-capacity 3D nylon membrane (comprising immobilized Protein A or Protein G) with an increased surface area, providing much higher protein-binding capabilities per ml of the membrane than per ml of resin.


References for antibody therapeutics and selected publications citing the use of In-Fusion Cloning for HTP antibody cloning

Chen, C. G. et al. One-step zero-background IgG reformatting of phage-displayed antibody fragments enabling rapid and high-throughput lead identification. Nuc. Acids Res. 42, e26 (2014).

Meng, W. et al. Efficient generation of monoclonal antibodies from single rhesus macaque antibody-secreting cells. mAbs 7, 707–718 (2015).

Spidel, J. L. et al. Rapid high-throughput cloning and stable expression of antibodies in HEK293 cells. J. Immunol. Methods 439, 50–58 (2016).

Thakur, A. et al. Bispecific antibody-based therapeutics: strengths and challenges. Blood Revs. 2, 004 (2018).

Wold, E. D. et al. Antibody therapeutics in oncology. Immunother. Open 2, 108 (2016).


Featured products

Cat. # Product Size License Quantity Details
638948 In-Fusion® Snap Assembly Master Mix 50 Rxns USD $665.00

In-Fusion Snap Assembly Master Mix is designed for fast, directional cloning of one or more fragments of DNA into any vector. This proprietary master mix fuses DNA fragments (e.g., PCR-generated sequences and linearized vectors) efficiently and precisely by recognizing a 15-bp overlap at their ends. This 15-bp overlap can be engineered into the primers designed for PCR amplification of the desired sequences. In Fusion Snap Assembly Master Mix offers high efficiency, even for applications that can be challenging, including the cloning of long fragments, short oligonucleotides, and multiple fragments.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data

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Performance comparison between In-Fusion Snap Assembly and NEBuilder HiFi using inverse PCR.

Performance comparison between In-Fusion Snap Assembly and NEBuilder HiFi using inverse PCR.

Performance comparison between In-Fusion Snap Assembly and NEBuilder HiFi using inverse PCR. A single 3.8-kb insert (Panel A) or a 34.2-kb adenovirus insert (Panel B) was cloned into a 2.7-kb vector which was linearized via inverse PCR. These cloning reactions were performed in triplicate with both In-Fusion Snap Assembly and NEBuilder HiFi. Primers were designed according to the manufacturers' specifications. After transformation and plating, 20 colonies from each replicate were analyzed by Sanger sequencing (for the 3.8-kb insert) or colony PCR (for the adenovirus insert) to determine the cloning accuracy. In-Fusion Snap Assembly yielded 2X more colonies than NEBuilder HiFi.

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638948: In-Fusion Snap Assembly Master Mix

638948: In-Fusion Snap Assembly Master Mix

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The In-Fusion cloning protocol

The In-Fusion cloning protocol

The In-Fusion cloning protocol.

639691 In-Fusion® HD EcoDry™ Cloning Kit 96 Rxns USD $1145.00

The In-Fusion HD EcoDry Cloning Kit enables high-efficiency, high-fidelity, directional cloning of one or more PCR fragments into any vector. This package includes a 96-well plate of In-Fusion HD EcoDry Mix, linearized pUC19 control vector, and a 2-kb control insert.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

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639691: In-Fusion HD EcoDry Cloning Kit

639691: In-Fusion HD EcoDry Cloning Kit
635731 Capturem™ Protein A 24-Well Plate 1 x 24-well plate USD $469.00

License Statement

ID Number  
273 This product is covered by U.S. Patent Nos. 9,459,188 and/or 10,207,229, exclusively licensed to Takara Bio USA, Inc.

Capturem Protein A 24-Well plates are single-use disposable plates that provide rapid, high-quality, small-scale purification of monoclonal and polyclonal antibodies. It combines the specific antibody-binding properties of Protein A with novel Capturem technology to provide high-capacity, membrane-based affinity purification. Typical yields for monoclonal antibodies are up to 400 µg/well with a sample load volume of up to 4.5 ml.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

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Capturem Protein A- or G-functionalized membranes

Capturem Protein A- or G-functionalized membranes

Antibody purification workflow for Capturem Protein A or G 96-well plates. Each well can be loaded with up to 850 µl of a diluted sample (antibody sample diluted from 1:1 to 1:20 with buffer). Following binding, the membrane is then washed and the bound antibiody eluted with the appropriate buffers. Note: Protocol time varies for each format. Check user manuals for specifics on protocol times.

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Capturem spin columns and filtration devices

Capturem spin columns and filtration devices

Available Capturem spin column and filtration device formats. Pictured from left to right: nanoprep, miniprep, 96-well, 24-well, maxiprep, and large-volume Capturem formats. Formats available vary for the different functionalized membranes. Check product pages for list of available formats.

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Load volumes and approximate yields for available Capturem formats by chemistry

Load volumes and approximate yields for available Capturem formats by chemistry

Load volumes and yields of Capturem purification formats.

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635731: Capturem Protein A 24-Well Plate

635731: Capturem Protein A 24-Well Plate
635732 Capturem™ Protein G 24-Well Plate 1 x 24-well plate USD $469.00

License Statement

ID Number  
273 This product is covered by U.S. Patent Nos. 9,459,188 and/or 10,207,229, exclusively licensed to Takara Bio USA, Inc.

Capturem Protein G 24-Well Plates are 24-well plates that provide rapid, high-quality, small-scale purification of monoclonal and polyclonal antibodies. It combines the specific antibody-binding properties of Protein G with novel Capturem technology to provide high-capacity, membrane-based affinity purification. Typical yields for monoclonal antibodies are up to 600 µg with a 4.5-ml sample capacity.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

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Capturem Protein A- or G-functionalized membranes

Capturem Protein A- or G-functionalized membranes

Antibody purification workflow for Capturem Protein A or G 96-well plates. Each well can be loaded with up to 850 µl of a diluted sample (antibody sample diluted from 1:1 to 1:20 with buffer). Following binding, the membrane is then washed and the bound antibiody eluted with the appropriate buffers. Note: Protocol time varies for each format. Check user manuals for specifics on protocol times.

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Capturem spin columns and filtration devices

Capturem spin columns and filtration devices

Available Capturem spin column and filtration device formats. Pictured from left to right: nanoprep, miniprep, 96-well, 24-well, maxiprep, and large-volume Capturem formats. Formats available vary for the different functionalized membranes. Check product pages for list of available formats.

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Load volumes and approximate yields for available Capturem formats by chemistry

Load volumes and approximate yields for available Capturem formats by chemistry

Load volumes and yields of Capturem purification formats.

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635732: Capturem Protein G 24-Well Plate

635732: Capturem Protein G 24-Well Plate

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