pINFUSE-hIgG1-Fc2 vector高效IgG Fc融合蛋白大量表达载体-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心

pINFUSE-hIgG1-Fc2 vector

Plasmid designed for the construction of Fc-Fusion proteins

-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心

Product information

Map图谱：

Content :

- 20 µg of pinfuse-higG1 -fc2 plasmid provided as lyophilized

DNA.

- 4 pouches of E. coli Fast-Media® Zeo (2 TB and 2 Agar)

sto rag e and stabi l i ty :

- Product is shipped at room temperature.

- Lyophilized DNA should be stored at -20˚C and is stable 3

months.

- Resuspended DNA should be stored at -20˚C and is stable up to 1

year.

- Store E. coli Fast-Media® Zeo at room temperature. Fast-Media®

pouches are stable 18 months when stored properly.

Qual i ty co ntro l :

- Plasmid construct has been confirmed by restriction analysis and

sequencing.

- Plasmid DNA was purified by ion exchange chromatography and

lyophilized.

General ProduCt use

pINFUSE-Fc is a family of plasmid developed to facilitate the

construction of Fc-fusion proteins by fusing the effector region

of a protein to the Fc region of an immunoglobulin G (IgG).

pINFUSE-Fc plasmids yield high levels of Fc-fusion proteins.

The level of expression is usually in the µg/mL range. They can

be transfected in a variety of mammalian cells, including

myeloma cell lines, CHO cells, monkey COS cells and human

embryonic kidney (HEK)293 cells, cells that are commonly used

in protein purification systems.

pINFUSE-Fc plasmids allow the secretion of Fc-Fusion proteins.

As Fc-Fusion proteins are secreted, they can be easily detected in

the supernatant of pINFUSE-Fc-transfected cells by SDS-PAGE.

Furthermore, functional domains can be identified by

immunoblotting and ligand blotting.

Fc-Fusion proteins can be easily purified by single-step protein

A or protein G affinity chromatography.BioVector provides pINFUSE-Fc vectors featuring Fc regions

containing introns from different species and isotypes. In

humans, there are four isotypes: IgG1, IgG2, IgG3 and IgG4. The

Fc region mediates effecto r functions, such as antibody -

dependent cellular cytotoxicity (ADCC) and complement -

dependent cytotoxicity (CDC). IgG isoforms exert different

levels o f effecto r functions increasing in the o rder o f

IgG4<IgG2<IgG1≤IgG3.

Plasmid features

• human g enomi c igG1 -fc (wi th i ntro ns ): The Fc region

comprises the CH2 and CH3 domains of the IgG heavy chain and

the hinge region. The hinge serves as a flexible spacer between the

two parts of the Fc-fusion protein, allowing each part of the

molecule to function independently. A short intron is present

between each region (one intron between the hinge and CH2 and one

intron between CH2 and CH3). The presence of introns is known to

enhance the level of gene expression as splicing is known to

promote rapid and efficient mRNA export1

.

Human IgG1 dispays high ADCC and CDC, and is the most

suitable fo r therapeutic use against pathogens and cancer

cells.

• hef1 -HtlV pro m is a composite promoter comprising the

Elongation Factor-1α (EF-1α) core promoter2 and the R segment

and part of the U5 sequence (R-U5’) of the Human T-Cell Leukemia

Virus (HTLV) Type 1 Long Terminal Repeat3

. The EF-1α promoter

exhibits a strong activity and yields long lasting expression of a

transgene in viv o. The R-U5’ has been coupled to the EF-1α core

promoter to enhance stability of RNA.

• il2 s s : The IL2 signal sequence contains 20 amino acids and

share common characteristics with signal peptides of other secretory

proteins. The intracellular cleavage of the IL2 signal peptide occurs

after Ser20 and leads to the secretion of the antigenic protein.

• mCs: The multiple cloning site contains several restriction sites

that are compatible with many other enzymes, thus facilitating

cloning.

• sV4 0 pan: the Simian Virus 40 late polyadenylation signal

enables efficient cleavage and polyadenylation reactions resulting

in high levels of steady-state mRNA4

.

• o ri : a minimal E. coli origin of replication to limit vector size,

but with the same activity as the longer Ori.

• CmV enh / hferl pro m: This composite promoter combines

the human cytomegalovirus immediate-early gene 1 enhancer and

the core promoter of the human ferritin light chain gene. This

ubiquitous promoter drives the expression of the Zeocin™-

resistance gene in mammalian cells.

• em2 KC is a bacterial promoter that enables the constitutive

expression of the antibiotic resistance gene in E. coli. EM2KC is

located within an intron and is spliced out in mammalian cells.

• Zeo : Resistance to Zeocin™ is conferred by the Sh ble gene from

Streptoalloteichus hindustanus The same resistance gene confers

selection in both mammalian cells and E. coli.

• βGl o pan: The human beta-globin 3’UTR and polyadenylation

sequence allows efficient arrest of the transgene transcription5

.