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Alzheimer's disease research

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Antibody engineering for Alzheimer's disease immunotherapies

Many neurodegenerative diseases are characterized by the abnormal accumulation of specific proteins in brain tissue. In Alzheimer's disease, these aggregates consist of amyloid plaques and neurofibrillary tangles (comprised of amyloid-beta [Aβ] and tau, respectively). As both Aβ and tau are neurotoxic, tend to correlate with cognitive decline, and induce inflammation, current models suggest that their aggregation triggers a cascade of downstream pathologies, drives neurodegeneration, and ultimately results in Alzheimer's disease (Bloom 2014; Laurent, Buée, and Blum 2018).

These protein aggregates, therefore, represent an attractive therapeutic target. A major push in clinical research has been in the development and testing of anti-Aβ and, more recently, anti-tau antibodies. While the field has experienced setbacks (Honig et al., 2018; Selkoe 2019), a growing number of antibodies have moved into clinical trials (reviewed in Cummings et al. 2018) and offer the hope that clearing these aggregates may arrest, and potentially reverse, the progression of this debilitating disease. To facilitate this work, we offer end-to-end antibody engineering solutions for simple, rapid, and high-throughput antibody generation and purification.

Highlighted products

SMARTer amplification of antibody variable regions

Our SMARTer RACE kits leverage our unique SMART (Switching Mechanism at the 5' end of RNA Template) technology to provide a true representation of the original heavy (H), kappa light chain  [L (κ)], or lambda light chain [L (λ)] mRNA transcripts from as little as 10 ng of total RNA (Figure 1). Unlike many other RACE-PCR methodologies, our SMARTer RACE technology captures the 5' end of transcripts and, coupled with the addition of the SMART sequence at the 5' end, provides full-length, uniform gene-body coverage. This optimized technology is robust, boasting low background and high amplification efficiency, and can be used even for samples contaminated with gDNA. Finally, to streamline your workflow, these kits include our proprietary In-Fusion Cloning technology (see below) to facilitate the rapid, highly accurate cloning of your RACE fragments.

Schematic for the SMARTer RACE kit workflow to amplify antibody variable regions

Figure 1. Overview of the experimental workflow for SMARTer 5' RACE of the antibody variable domain. 

Efficient, high-throughput generation of antibody constructs

Our In-Fusion HD Cloning Plus technology (Figure 2) enhances antibody discovery and engineering workflows by allowing the rapid generation and screening of functional clones. Screening for individual clones is usually one of the most time-consuming steps of antibody engineering workflows. However, unlike traditional ligation-based methods, In-Fusion Cloning is highly efficient, has an extremely low background, and boasts a >95% success rate for single-insert cloning. Our In-Fusion technology is also fast (15 minutes) and seamless, allowing the directional cloning of up to several PCR fragments into any destination vector in a single reaction. Due to these benefits, our In-Fusion technology has powered multiple high-throughput antibody cloning workflows (Chen et al. 2014; Meng et al. 2015; Spidel et al. 2016; Rudkin et al. 2018).

In-fusion cloning workflow overview.

Figure 2. Overview of the In-Fusion Cloning workflow.

After antibodies have been generated from your expression construct, our next-generation Capturem purification technology enables rapid, high-quality, and high-throughput antibody purification within 5–15 minutes. Capturem columns and plates are resin-free and leverage a high-capacity nylon-membrane-based system (Figure 3) that contains immobilized Protein A or G (Figure 3), allowing direct antibody purification from complex starting samples (such as hybridoma supernatants and serum). These technologies are available in a variety of formats (minipreps, maxipreps, 24-well plates, and 96-well plates) and are ideal for rapid, high-throughput screening of your antibody clones.

Image detailing the interior of a Capturem high-capacity nylon membrane.

Figure 3. Image detailing the interior of a Capturem high-capacity membrane.


References for AD and the development of Αβ and tau-targeted antibodies

Bloom, G. S. Amyloid-β and tau. JAMA Neurol. 71, 505 (2014).

Cummings, J., Lee, G., Ritter, A. & Zhong, K. Alzheimer's disease drug development pipeline: 2018. Alzheimer's Dement. (New York, N. Y.) 4, 195–214 (2018).

Honig, L. S. et al. Trial of solanezumab for mild dementia due to Alzheimer's disease. N. Engl. J. Med. 378, 321–330 (2018).

Laurent, C., Buée, L. & Blum, D. Tau and neuroinflammation: What impact for Alzheimer's disease and tauopathies? Biomed. J. 41, 21–33 (2018).

Selkoe, D. J. Alzheimer disease and aducanumab: adjusting our approach. Nat. Rev. Neurol. 15, 365–366 (2019).

References citing the use of In-Fusion HD Cloning for high-throughput antibody cloning

Chen, C.-G., Fabri, L. J., Wilson, M. J. & Panousis, C. One-step zero-background IgG reformatting of phage-displayed antibody fragments enabling rapid and high-throughput lead identification. Nucleic Acids Res. 42, e26–e26 (2014).

Meng, W. et al. Efficient generation of monoclonal antibodies from single rhesus macaque antibody secreting cells. MAbs 7, 707–18 (2015).

Rudkin, F. M. et al. Single human B cell-derived monoclonal anti-Candida antibodies enhance phagocytosis and protect against disseminated candidiasis. Nat. Commun. 9, 5288 (2018).

Spidel, J. L., Vaessen, B., Chan, Y. Y., Grasso, L. & Kline, J. B. Rapid high-throughput cloning and stable expression of antibodies in HEK293 cells. J. Immunol. Methods 439, 50–58 (2016).


Featured products

Cat. # Product Size License Quantity Details
634858 SMARTer® RACE 5’/3’ Kit 10 Rxns USD $801.00

The SMARTer RACE 5'/3' Kit allows the synthesis of first-strand cDNA from poly A+ or total RNA via SMART (Switching Mechanism At 5' End of RNA Template) technology, and facilitates the performance of 5'- and 3'-RACE (Rapid Amplification of cDNA Ends) PCR with the kit's Universal Primer Mix. Our carefully designed, specially-modified SMARTer Oligo preferentially captures the 5' ends of the cDNA during cDNA synthesis. Using this SMARTer Oligo, our procedure enriches cDNA pools for 5' sequences, thus increasing the likelihood you will amplify the entire sequence of your gene.

RACE PCR products are amplified with the provided SeqAmp DNA Polymerase, and cloned into the linearized pRACE vector with In-Fusion HD Cloning. The SMARTer RACE 5'/3' Kit has been improved to accommodate larger RNA input volumes and perform better on challenging targets than the original SMARTer RACE cDNA Amplification Kit. The In-Fusion HD Cloning Kit, NucleoSpin Gel and PCR Clean-Up Kit, and Stellar Competent Cells are included for your convenience in cloning RACE products. Gene-specific RACE primers are supplied by the user.

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.

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Overview of the SMARTer RACE 5'/3' Kit workflow

Overview of the SMARTer RACE 5'/3' Kit workflow
Overview of the SMARTer RACE 5'/3' Kit workflow. Each kit is a complete system, containing the reagents required to recover cloned RACE fragments on the second day.

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634858: SMARTer RACE 5'/3' Kit

634858: SMARTer RACE 5'/3' Kit
635731 Capturem™ Protein A 24-Well Plate 1 x 24-well plate USD $502.00

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.

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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
635745 Capturem™ Protein G 24-Well Plate - HC 1 x 24-Well Plate USD $502.00

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.

The Capturem Protein G 24-Well Plate - HC is a high-capacity, single-use disposable plate for the simple, rapid purification of antibodies. Up to 4.3 ml of sample can be loaded per well. This plate is suitable for purification of antibodies from animal serum, ascites fluid, cell culture supernatant of hybridoma cell lines, and other sources.

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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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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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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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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635745: Capturem Protein G 24-Well Plate - HC

635745: Capturem Protein G 24-Well Plate - HC
638920 In-Fusion® HD Cloning Plus 96 Rxns USD $1790.00

We are in the process of discontinuing In-Fusion HD. We recommend you instead order the In-Fusion Snap Assembly Starter Bundle (10-rxn) or the In-Fusion Snap Assembly Value Bundle (50-rxn).

The In-Fusion HD Cloning Plus kit enables high-efficiency, high-fidelity, directional cloning of one or more PCR fragments into any vector. In addition to the cloning kit, this package includes: a NucleoSpin 96 PCR Clean-Up kit, which can be used for spin-column purification of PCR products; Stellar Competent Cells, an E. coli HST08 strain that provides high transformation efficiency paired with blue-white screening capability when used with pUC plasmid vectors; and CloneAmp HiFi PCR Premix, a convenient high-fidelity PCR 2X master mix that provides exceptionally accurate and efficient DNA amplification.

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.

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The In-Fusion HD Cloning System has been optimized for cloning large fragments

The In-Fusion HD Cloning System has been optimized for cloning large fragments
The In-Fusion HD Cloning System has been optimized for cloning large fragments. Ten out of ten colonies contain the correct insert (100% efficiency) when cloning fragments as large as 15 kb (results confirmed by colony PCR screening).

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The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction
The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.

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The In-Fusion cloning protocol

The In-Fusion cloning protocol
The In-Fusion cloning protocol.

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638920: In-Fusion HD Cloning Plus

638920: In-Fusion HD Cloning Plus
639691 In-Fusion® HD EcoDry™ Cloning Kit 96 Rxns USD $1225.15

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

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Takara Bio USA, Inc. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, TBUSA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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