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  • Characterizing the viral genome and host response
  • Identifying and cloning protein targets
  • Expressing and purifying protein targets
  • Immunizing mice and optimizing vaccines

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Home › Applications › mRNA and protein therapeutics › Expressing and purifying protein targets

mRNA and protein therapeutics

  • Characterizing the viral genome and host response
  • Identifying and cloning protein targets
  • Expressing and purifying protein targets
  • Immunizing mice and optimizing vaccines
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Expressing and purifying vaccine targets

Recombinant subunit protein vaccines are safe, potent, and well-characterized options that provide broad protection against multiple serotypes with low reactogenicity. Researchers working with recombinant protein-based subunit vaccines need to express their vaccine candidate proteins at high levels with a small, well-tolerated tag such as the 6xHis-tag to allow downstream purification. Choosing an efficient system for the optimal expression and efficient purification of the vaccine candidates is essential to speed up the vaccine development process.

Our BakPAK Baculovirus Expression System is ideal for obtaining high recombinant protein yields compared to other expression systems. It is also optimal for expressing proteins that are difficult to express in bacteria due to size, complexity, or post-translational modifications. BakPak technology is easy to scale up, has high protein expression levels, and is nonpathogenic to humans. The post-translational processing and folding of recombinant proteins produced in insect cells more closely resemble mammalian processes.

Our BacPAK Baculovirus Rapid Titer Kit uses a standard immunological assay to accurately determine baculovirus titers within 48 hours, while plaque and end-point dilution assays typically require 5–10 days, and is compatible with all commonly available (AcMNPV-based) baculovirus expression systems.

Once expressed from these purified constructs, his-tagged vaccine candidate proteins can be rapidly purified using our performance-validated cobalt and nickel resins or our revolutionary Capturem membrane-based purification technology. Capturem technology utilizes high-capacity nylon membranes immobilized with Ni-NTA, allowing rapid and easy his-tagged protein purification in under 30 minutes. In contrast, typical resin-based purification methods using immobilized metal affinity chromatography (IMAC) columns require several hours to complete due to long column equilibration/binding times and slow diffusion of large macromolecules through the resin bed.

Learn about the many ways Capturem technology can help speed up your research.

Benefits

  • Fast, convenient workflow—5- to 30-min room-temperature protocol using convenient spin columns/plates/filtration devices featuring Capturem nickel-functionalized membranes
  • High purity—small bed volumes trap fewer contaminants
  • Compatible with a wide range of additives—including EDTA, DTT, BME, glycerol, TCEP, etc. (see compatibility table)
  • Versatile—easy purification from mammalian/bacterial cell lysates and supernatants

 


Numerous laboratories have already used our TALON resin and Capturem technology for their vaccine development research:

Title Link Product
Protein purification
SARS-CoV-2
A rapid, point of care red blood cell agglutination assay for detecting antibodies against SARS-CoV-2 Capturem His-Tagged Purification Miniprep Kit
Dynamics of neutralizing antibody titers in the months after SARS-CoV-2 infection TALON resin
Functional SARS-CoV-2-specific immune memory persists after mild COVID-19
Pre-existing and de novo humoral immunity to SARS-CoV-2 in humans
SARS-CoV-2 and bat RaTG13 spike glycoprotein structures inform on virus evolution and furin-cleavage effects
Ultra-sensitive high-resolution profiling of anti-SARS-CoV-2 antibodies for detecting early seroconversion in COVID-19 patients
Other viruses
Recombinant adenovirus carrying a core neutralizing epitope of porcine epidemic diarrhea virus Capturem His-Tagged Purification Miniprep Kit
Generation of the recombinant antigen LC3 from Entamoeba histolytica linked to mutated exotoxin A (PEΔIII)

Featured products

Cat. # Product Size Price License Quantity Details
631402 BacPAK™ Baculovirus Expression System Each USD $1054.00

Complete kit for expressing recombinant proteins at high levels. The BacPAK System uses the baculovirus Autographa californica nuclear polyhedrosis virus (AcMNPV) to produce target proteins in insect cells. More than 90% of the viruses produced by the transfected cells carry the target protein. The expressed recombinant protein is similar in structure, biological activity, and immunological reactivity to the naturally occurring protein because insect host cells provide post-translational processing similar to that of mammalian cells.

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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BacPAK6 Viral DNA map

BacPAK6 Viral DNA map
BacPAK6 Viral DNA map.

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Protein production from recombinant viruses generated using the BacPAK Baculovirus Expression System

Protein production from recombinant viruses generated using the BacPAK Baculovirus Expression System
Protein production from recombinant viruses generated using the BacPAK Baculovirus Expression System. Recombinant viruses were obtained by cotransfection of transfer vectors with BacPAK6 Viral DNA (Bsu36 I digest), followed by amplification in Sf21 cells. The SDS PAGE analysis of cellular lysates was performed 48 hr after infection of the Sf21 cultures. Lane 1: uninfected Sf21 cells. Lane 2: Sf21 cells infected with wild-type AcMNPV virus. Lane 3: Sf21 cells infected with nonrecombinant BacPAK6 virus. Lane 4: Sf21 cells infected with BacPAK8-GUS recombinant virus. Lane 5: purified CAT protein. Lane 6: Sf21 cells infected with BacPAK9-CAT recombinant virus. Lane M: molecular weight marker.
631406 BacPAK™ Baculovirus Rapid Titer Kit 5 Assays USD $574.00

License Statement

ID Number  
330 This product is the subject of a technology license agreement for internal research use only. Use of this product other than for research use may require additional licenses. Information on license restrictions or for uses other than research may be obtained by contacting licensing@takarabio.com.

The BacPAK Baculovirus Rapid Titer Kit enables you to determine infectious baculovirus titers based on the expression of baculoviral gp64. The immunological staining protocol can be completed in 2 days.

Cat. # 631406 includes Goat Anti-mouse Antibody/HRP Conjugate, Normal Goat Serum, Mouse gp64 Antibody, Blue Peroxidase Substrate, Methyl Cellulose Overlay, and Resealable Plastic Bags.

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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Identification of infection foci in a lawn of Sf21 cells, using the BacPAK Baculovirus Rapid Titer Kit

Identification of infection foci in a lawn of Sf21 cells, using the BacPAK Baculovirus Rapid Titer Kit
Identification of infection foci in a lawn of Sf21 cells, using the BacPAK Baculovirus Rapid Titer Kit. Cells were immunostained with the gp64 primary antibody and an HRP-conjugated secondary antibody, and visualized by light microscopy. Panel A. Former protocol, with formyl-buffered acetone fixation. Panel B. Improved protocol, with 4% paraformaldehyde fixation.

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The In-Fusion Ready BacPAK Vector Set and Baculovirus Expression System

The In-Fusion Ready BacPAK Vector Set and Baculovirus Expression System
The In-Fusion Ready BacPAK Vector Set and Baculovirus Expression System. A PCR fragment containing your gene of interest is simultaneously and directly cloned into the In-Fusion Ready BacPAK Vector pair to generate N- and C-terminal-tagged constructs. Verified clones are then cotransfected with BacPAK6 viral DNA into insect cells to initiate recombination via the AcMNPV sequences, and replication of recombinant virus. Following virus amplification, infection of insect cell cultures produces recombinant protein, which is then purified by TALON technology.

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Insect cells infected with a recombinant baculovirus express the Aequorea coerulescens green fluorescent protein (AcGFP1)

Insect cells infected with a recombinant baculovirus express the Aequorea coerulescens green fluorescent protein (AcGFP1)
Insect cells infected with a recombinant baculovirus express the Aequorea coerulescens green fluorescent protein (AcGFP1). An In-Fusion Ready BacPAK vector was used to generate a recombinant baculovirus harboring an N-terminal tagged expression construct for the Living Colors fluorescent protein, AcGFP1. Sf9 cells infected with the virus expressed high levels of AcGFP1 and became highly fluorescent. Analysis by flow cytometry revealed that the mean fluorescence intensity of the infected cells was approximately 440-fold greater than that of the uninfected control.

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Insect cells express N- and C-tagged Aequorea coerulescens green fluorescent protein (AcGFP1)

Insect cells express N- and C-tagged Aequorea coerulescens green fluorescent protein (AcGFP1)
Insect cells express N- and C-tagged Aequorea coerulescens green fluorescent protein (AcGFP1). In-Fusion Ready BacPAK vectors were used to generate recombinant baculoviruses harboring N- or C-terminal-tagged AcGFP1 expression constructs. Insect cells (Sf9) infected with either virus express AcGFP1 and thus appear green when viewed by fluorescence microscopy. Panel A. BacPAK-Nterm-6xHN-AcGFP1. Panel B. BacPAK-Cterm- 6xHN-AcGFP1

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Flow chart of the two-day BacPAK Baculovirus Rapid Titer Kit procedure

Flow chart of the two-day BacPAK Baculovirus Rapid Titer Kit procedure
Flow chart of the two-day BacPAK Baculovirus Rapid Titer Kit procedure.

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631406: BacPAK Baculovirus Rapid Titer Kit

631406: BacPAK Baculovirus Rapid Titer Kit
635730 Capturem™ His-Tagged Purification 24-Well Plate 1 x 24-well plate USD $480.00

License Statement

ID Number  
273 This Product is protected by one or more patents from the family comprising: US9459188, US10207229 and any corresponding patents, divisionals, continuations, patent applications and foreign filings sharing priority with the same family.
347 This product is protected by U.S. Patents 9,895,665, 10,195,569 and foreign counterparts and/or additional U.S. and foreign patents pending. For further license information, please contact a Takara Bio USA licensing representative by email at licensing@takarabio.com.

Capturem His-tagged 24-Well Plates are 24-well plates that provide rapid, high-quality, small-scale purification of his-tagged proteins. These plates combine Ni-NTA chemistry with novel Capturem technology to provide high-capacity, membrane-based affinity purification. Typical yields for his-tagged proteins are up to 800 µg per well with a 4.5-ml sample capacity per well.

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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635730: Capturem His-Tagged Purification 24-Well Plate

635730: Capturem His-Tagged Purification 24-Well Plate

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Capturem nickel-functionalized membranes

Capturem nickel-functionalized membranes

Purification workflow for Capturem His-Tagged Purification 96-Well Plate. The his-tagged protein of interest is first bound to the membrane, and then washed and 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.

635660 His60 Ni Superflow Resin 25 mL USD $331.00

A high-capacity immobilized metal affinity chromatography resin for the efficient purification of recombinant his-tagged proteins under native or denaturing conditions.

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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Nickel resin—His60 for his-tag purification of proteins

Nickel resin—His60 for his-tag purification of proteins
Nickel resin—His60 for his-tag purification of proteins.

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635660: His60 Ni Superflow Resin

635660: His60 Ni Superflow Resin
635714 Capturem™ His-Tagged Purification 96-Well Plate 1 x 96-well plate USD $484.00

License Statement

ID Number  
273 This Product is protected by one or more patents from the family comprising: US9459188, US10207229 and any corresponding patents, divisionals, continuations, patent applications and foreign filings sharing priority with the same family.
347 This product is protected by U.S. Patents 9,895,665, 10,195,569 and foreign counterparts and/or additional U.S. and foreign patents pending. For further license information, please contact a Takara Bio USA licensing representative by email at licensing@takarabio.com.

Capturem His-Tagged Purification 96 is a single-use, disposable 96-well plate for simple, rapid purification of his-tagged proteins from up to 1 ml of clarified lysate per well. This plate is suitable for use under native or denaturing conditions, in the presence of additives such as DTT (up to 10 mM), βME (up to 30 mM), TCEP (up to 5 mM), EDTA (up to 10 mM), or glycerol. Capturem His-Tagged Purification 96 is compatible with automated liquid handlers equipped with vacuum filtration or centrifuges equipped with appropriate plate holders. Each well requires a minimum elution volume of 100 µl.

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

Back

635714: Capturem His-Tagged Purification 96-Well Plate

635714: Capturem His-Tagged Purification 96-Well Plate

Back

Capturem nickel-functionalized membranes

Capturem nickel-functionalized membranes

Purification workflow for Capturem His-Tagged Purification 96-Well Plate. The his-tagged protein of interest is first bound to the membrane, and then washed and eluted with the appropriate buffers. Note: protocol time varies for each format. Check user manuals for specifics on protocol times.

Back

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.

Back

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.


Capturem membrane technology

Capturem technology

There is a constant need for faster, more efficient antibody and protein purification processes at any scale. Capturem membrane technology allows for purification directly from complex matrices, such as cell supernatants, in minutes. This technology also provides highly purified and concentrated antibodies and his-tagged proteins, even from samples containing additives not compatible with other purification technologies.

Antibody purification His-tagged purification

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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, Takara Bio USA 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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  • Single-cell NGS automation
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  • Great value master mixes
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