pTRIOZ-hIgG4 (S228P)

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pTRIOZ-hIgG4 (S228P)

Plasmid for production of human IgG4 kappa mAbs (without signal sequence)

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20 µg

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  • Specifications
  • Contents
  • Details

pTRIOZ General plasmid map

High-yield production of recombinant human IgG4 kappa mAbs 

pTRIOZ-hIgG4 (S228P)is designed for high-yield production of whole monoclonal antibodies (mAbs) with the use of a single plasmid. It consists of three cassettes encoding for the expression of the mAb heavy chain, light chain, and antibiotic selection with Zeocin®

pTRIOZ-hIgG4 (S228P) expresses the constant regions of:

  • The human IgG4 heavy chain with the S228P mutation, ensuring the stabilization of the disulfide bonds in the core-hinge region of IgG4 molecules to reduce Fab-arm exchange. 
  • The human kappa light chain

Upstream of both of these regions are unique multiple cloning sites (MCS) that enable the insertion of the variable regions of any given mAb.

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Heavy chain cassette:

  • Composite promoter: Aldehyde dehydrogenase ( AldA ) enhancer/ Human ferritin heavy chain gene ( hFerH )
  • Cloning sites: 5’- AgeI, MluI, EcoRV, and NheI -3’
  • Constant region: Human IgG4 (S228P)

Light chain cassette:

  • Composite promoter: Human cytomegalovirus immediate-early gene 1 ( hCMV ) enhancer / Human ferritin light chain gene ( hFerL )
  • Cloning sites: 5’- SgrAI, AscI, PmeI, NcoI, and BsiWI -3’.
  • Constant region: Human kappa 

Antibiotic resistance: Zeocin® (selectable in both bacterial and mammalian cells)

Quality control:

  • Plasmid construct has been confirmed using DNA sequencing and restriction analysis
  • Purified by ion exchange chromatography
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  • 20 µg of lyophilized pTRIOZ-hIgG4 (S228P) plasmid DNA
  • 1 ml of Zeocin® (100 mg/ml)


Room temperature Product is shipped at room temperature

Store Upon receipt product should be stored at -20°C

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

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Cassette 1: mAb Heavy chain

  • AldA enh/ hFerH: This composite promoter combines the human aldehyde dehydrogenase (aldA) enhancer and the core promoter of the human ferritin heavy chain gene. 
  • MCS1: To facilitate cloning of the variable heavy (VH) chain, the multiple cloning site contains restriction sites that are compatible with many different enzymes, 
  • hIgG4: The constant region of the human immunoglobulin IgG4 heavy chain with the core-hinge stabilizing S228P mutation.
  • βGlo pAn: The human beta-globin 3’UTR and polyadenylation sequence allow the efficient arrest of the transgene transcription.


Cassette 2: mAb Light chain

  • hCMV enh / hFerL prom: This composite promoter combines the human cytomegalovirus (CMV) immediate-early gene 1 enhancer and the core promoter of the human ferritin light chain gene. 
  • MCS2: To facilitate cloning of the variable light (VL) chain, the multiple cloning site contains restriction sites that are compatible with many different enzymes, 
  • Human κ light chain: The constant region of the human kappa light chain
  • SV40 pAn: The Simian Virus 40 late polyadenylation signal enables efficient cleavage and polyadenylation reactions resulting in high levels of steady-state mRNA


Cassette 3: Zeocin™ selection

  • mCMV/hEF1-HTLV prom: This composite promoter combines mouse cytomegalovirus (mCMV) immediate-early gene 1 enhancer, the elongation Factor-1α (EF-1α) core promoter, as well as the R  segment and part of the U5 sequence (R-U5’) of the Human T-Cell Leukemia Virus (HTLV) type 1 long terminal repeat. The EF-1α promoter exhibits a strong activity and yields a long-lasting expression of a transgene in vivo. The R-U5’ has been coupled to the EF-1α core promoter to enhance the stability of RNA.
  • EM7 prom: This is a bacterial promoter that enables the constitutive expression of the antibiotic resistance gene in E. coli. EM7 is located within an intron and is spliced out in mammalian cells.
  • Sh Ble gene: Resistance to Zeocin™ is conferred by the Sh ble gene from Streptoalloteichus hindustanus. The same gene confers resistance in both mammalian cells and E. coli.
  • hEF-1alpha pAn: This provides a strong polyadenylation signal. InvivoGen uses a sequence that starts after the stop codon of the EF1 cDNA and finishes after a bent structure rich in GT.


General features: pTRIOZ

  • 5’ UTR: The 5’ UTR enhances mRNA stability and protein translation.
  • Ori: A minimal E. coli origin of replication.
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