Providing functional, high-purity recombinant proteins—including membrane proteins and nanodiscs—to overcome bottlenecks in drug screening and target validation.
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Stable expression over 15 generations with rapid cell line development in just 3 months. Supports adherent and suspension cell lines, offering MCB, WCB, and PCB establishment.
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Cat. No. :
VET1375
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Product Information
Cat. No.
VET1375
Host Cell
Mammalian cells, Escherichia coli
Promoter
CMV-IE
Resistance
Ampicillin
Vector Type
Expression Vector
Vector Size
4758 bp
Quick Inquiry
Background
Case Study
Applications
Publications
Q & A
Customer Reviews
The pCAGGS Vector is a Mammalian Expression Vector sourced and authored by De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R. This vector, which has a length of 4801 base pairs, is resistant to the antibiotic ampicillin, a feature that plays a significant role in the selection process of successfully transformed cells in bacterial culture.
The pCAGGS Vector is renowned for its high efficiency in expressing genes under the regulation of the AG promoter and the human CMV-IE enhancer in an array of mammalian cells. This is made possible by the unique composition of the AG promoter sequence. The AG promoter combines elements from the chicken β-actin promoter, including the first exon as well as part of the first intron which appears to possess strong enhancer-like activity. In addition to these components, the AG promoter is also linked to a section of the rabbit β-globin fragment. This segment encompasses a 3' part of the second intron, which includes a branch point that is crucial for normal splicing reactions, and a 5' part of the third exon.
Case Study 1
Here, researchers report an electrochemical system for the detection of specific antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) proteins in patient serum samples following coronavirus disease 2019 (COVID-19). To this end, the recombinant SARS-CoV-2 spike protein (SCoV2-rS) was covalently immobilised on the surface of the gold electrode pre-modified with mixed self-assembled monolayer (SAMmix) consisting of 11-mercaptoundecanoic acid and 6-mercapto-1-hexanol. The affinity interaction of SCoV2-rS with specific antibodies against this protein (anti-rS) was detected using two electrochemical methods: cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The developed electrochemical immunosensor is suitable for confirming COVID-19 infection or the immune response after human vaccination.
Figure 1. Schematic representation of experimental stages. pCAGGS (Creative Biogene, cat. no. VET1375) is involved in the preparation of the SCoV2-rS protein. (Liustrovaite V, et al., 2022)
The SCoV2-rS protein is produced in mammalian Chinese hamster ovary (CHO) cells as a secreted trimeric protein. To match the native conformation locked in the prefusion state, the gene encoding the extracellular domain of SARS-CoV-2 Spike (SCoV2-S), including amino acids (aa) 1-1208, was chemically synthesized. The synthesized gene was then cloned into the mammalian expression vector pCAGGS (Creative Biogene, catalog number VET1375) by introducing NotI and XhoI restriction sites at the 5' and 3' gene ends, respectively. The entire expression construct includes full-length SCoV2-S ectodomain (aa 1–1208) w/o transmembrane and cytoplasmic aa, furin cleavage site 'RRAR' mutated to 'GSAS', C-terminal GSN4 trimerization motif fused to protein sequence, followed by thrombin cleavage site. Two mutations (K986P and V987P) were also introduced into the SCoV2-S sequence to stabilize the trimer in the prefusion conformation.
Case Study 2
Serological diagnosis and vaccination effectiveness assessment of coronavirus disease 2019 (COVID-19) is determined by the presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific antibodies. Here, researchers present an electrochemical-based biosensing technology for the detection of SARS-CoV-2 protein-specific antibodies. Recombinant SARS-CoV-2 spike protein (rSpike) was immobilized on the surface of a self-assembled monolayer (SAM)-modified gold electrode. The modified electrode was used as a sensitive element for the detection of polyclonal mouse antibodies against rSpike (anti-rSpike). The results demonstrate that the impedance measurement method and refined experimental conditions are suitable for further development of electrochemical biosensors for serological diagnosis of COVID-19 and/or confirmation of successful vaccination against SARS-CoV-2.
Figure 2. Schematic representation of experimental stages. pCAGGS (Creative Biogene, cat. no. VET1375) is involved in the preparation of the rSpike protein. (Drobysh M, et al., 2022)
Hamster CHO cells were used to secrete rSpike protein. This gene encodes the SARS-CoV-2 Spike extracellular domain, including amino acids (aa) 1–1208. The gene was integrated into the expression vector pCAGGS (Creative Biogene, catalog number VET1375) through the restriction sites NotI and XhoI added correspondingly at the 5' end and 3' end of the gene. These expression constructs contain the following: (i) full-length rSpike ectodomain (aa 1–1208) without transmembrane and cytoplasmic aa, (ii) furin cleavage site 'RRAR' mutated to 'GSAS', (iii) C-terminal GSN4 trimerization motif fused to the protein sequence, (iv) thrombin cleavage site. Two mutations (K986P and V987P) were introduced into the rSpike sequence to stabilize the trimer in the prefusion conformation.
Background
This study presents an electrochemical biosensing approach to detect antibodies specific to SARS-CoV-2. Recombinant SARS-CoV-2 spike protein (rSpike) was covalently immobilised on a gold electrode modified with an 11-mercaptoundecanoic acid self-assembled monolayer. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) monitored each surface modification step, including antibody binding. EIS revealed increasing charge-transfer resistance after rSpike immobilisation and anti-rSpike immunocomplex formation. The findings support further development of impedimetric immunosensors for COVID-19 serological diagnosis and vaccination evaluation. Key Findings
The study demonstrated that recombinant SARS-CoV-2 spike protein (rSpike) can be covalently immobilised onto an 11-MUA self-assembled monolayer on a gold electrode via EDC/NHS activation. Cyclic voltammetry showed that the 11-MUA SAM blocked electron transfer at the Au electrode, while EIS revealed stepwise changes: charge-transfer resistance (Rct) increased after SAM formation (4.58 kΩ·cm²), decreased after EDC/NHS activation (2.38 kΩ·cm²), increased after rSpike immobilisation (5.45 kΩ·cm²), and further increased after anti-rSpike antibody binding (6.41 kΩ·cm²). Bode plots showed impedance values at 0.1 Hz increasing from 10.6 kΩ·cm² for Au/SAM to 12.6 kΩ·cm² for Au/SAM/rSpike and 14.5 kΩ·cm² for Au/SAM/rSpike/anti-rSpike, compared with 6.2 kΩ·cm² for activated Au/SAM/EDC-NHS. The results indicate that EIS in Nyquist/Bode formats is sensitive to antigen-antibody complex formation and suitable for developing electrochemical immunosensors for COVID-19 serological diagnosis and vaccination assessment. Contribution
Creative Biogene provided the pCAGGS expression vector (cat. no. VET1375), which was used to construct the recombinant SARS-CoV-2 spike ectodomain expression plasmid. This vector enabled rSpike protein production in CHO cells, supporting the study's antigen preparation for electrochemical antibody detection.
Case Study 4
Calibration curves obtained from CV (a) and EIS fitted Rct (b), EIS, maximum of Z″ (c).
Background
This study reports an electrochemical immunosensor for detecting antibodies against SARS-CoV-2 spike protein. Recombinant SARS-CoV-2 spike protein was covalently immobilized on a gold electrode modified with a mixed self-assembled monolayer of 11-mercaptoundecanoic acid and 6-mercapto-1-hexanol. Affinity binding with specific anti-spike antibodies was monitored by cyclic voltammetry and electrochemical impedance spectroscopy. The sensor achieved detection limits of 2.53 nM and 1.99 nM for CV and EIS, respectively. The platform shows promise for serological COVID-19 diagnosis and vaccination immune response assessment. Key Findings
An electrochemical immunosensor was developed by covalently immobilizing recombinant SARS-CoV-2 spike protein (SCoV2-rS) onto a gold electrode pre-modified with a mixed self-assembled monolayer (SAMmix) of 11-mercaptoundecanoic acid and 6-mercapto-1-hexanol. Each modification step was characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Formation of SAMmix increased charge transfer resistance (Rct) from 141 to 884 Ω·cm2, EDC-NHS activation decreased it to 300 Ω·cm2, and SCoV2-rS immobilization increased it to 1280 Ω·cm2. Sequential incubation with anti-rS antibodies decreased CV peak currents and increased EIS semicircles, indicating immune complex formation. The sensor detected anti-rS with detection limits of 2.53 nM by CV and 1.99 nM by EIS using the maximum Z″ signal. Saturation occurred at 90–150 nM. The platform is suitable for serological COVID-19 diagnosis and monitoring vaccine-induced immunity. Contribution
Creative Biogene supplied the pCAGGS mammalian expression vector (cat. no. VET1375) used to clone the SARS-CoV-2 spike ectodomain construct. This vector enabled expression of recombinant trimeric spike protein in CHO cells for immunosensor development. The vector was crucial for producing the recombinant SCoV2-rS antigen used in the electrochemical immunosensor.
The pCAGGS vector is broadly utilized in the field of molecular biology for different applications, particularly in gene expression studies. Here are some applications of the pCAGGS vector:
Cloning and Expression of Genes: The pCAGGS vector is often used as a carrier or vehicle to insert a specific gene into a host cell to produce the protein coded by the inserted gene. It has a high expression level in mammalian cells, which makes it an ideal vector for this purpose.
Generation of Recombinant Proteins: The pCAGGS vector can be used to generate recombinant proteins. These proteins might have potential commercial or therapeutic uses.
Studying Gene Function: By inserting a specific gene into the pCAGGS vector and then introducing it into cells, scientists can study the function and behavior of this gene. This application is crucial for genetic research, medical research, and drug discovery.
Vaccine Development: The pCAGGS vector is also heavily used in the development of viral vaccines. It is used in the generation of viral proteins that can train the immune system to recognize and neutralize viruses.
Genetically Modified Organisms: The pCAGGS vector can be used to insert specific genes into plants or animals to create genetically modified organisms (GMOs). These GMOs can have desirable characteristics such as increased yield or resistance to pests.
Publications
Liustrovaite V, Drobysh M, Rucinskiene A, et al. Towards an electrochemical immunosensor for the detection of antibodies against SARS-CoV-2 spike protein[J]. Journal of The Electrochemical Society, 2022, 169(3): 037523.
Drobysh M, Liustrovaite V, Baradoke A, et al. Electrochemical determination of interaction between SARS-CoV-2 spike protein and specific antibodies[J]. International journal of molecular sciences, 2022, 23(12): 6768.
Customer Q&As
What is the pCAGGS vector?
A: The pCAGGS vector is used for the highly efficient expression of genes under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.
What is the composition of the AG promoter sequence?
A: The AG promoter sequence consists of the chicken β-actin promoter, the first exon, part of the first intron (with strong enhancer-like activity), a rabbit β-globin fragment (3' part of the second intron and 5' part of the third exon).
Why is it advisable to use lacIq strains when cloning a fragment downstream from the lac promoter?
A: Using lacIq strains helps prevent fortuitous expression of a possibly harmful polypeptide.
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Customer Reviews
Flexibility
The pCAGGS vector deserves praise for its flexibility and broad host range thus allowing it to be used in different kinds of mammalian cells.
French
Consistency
The pCAGGS vector is definitely a reliable tool in the field of molecular biology. The pCAGGS vector is known for its consistency and reliable results.
Canada
Easy to use
The pCAGGS vector is an excellent tool for cloning and expressing genes of interest. It's easy to use, versatile, and efficient in transfecting a wide variety of cell types.
French
Reliable vector
I have consistently had positive experiences with the pCAGGS vector. Its high efficiency of gene transfer and expression, combined with its robustness in multiple mammalian cell types, makes it a reliable vector for research purposes.
United States
A broad tropism for cell types
The pCAGGS vector has a broad tropism for cell types, even difficult-to-transfect cell types, making it reliable and practical for genetic research.
United Kingdom
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Figure 1. Schematic representation of experimental stages. pCAGGS (Creative Biogene, cat. no. VET1375) is involved in the preparation of the SCoV2-rS protein. (Liustrovaite V, et al., 2022)
Figure 2. Schematic representation of experimental stages. pCAGGS (Creative Biogene, cat. no. VET1375) is involved in the preparation of the rSpike protein. (Drobysh M, et al., 2022)
Bode plots of differently modified Au electrode
