pFUSE-Lucia-CHIg-mG1

Heavy chain constant region expression plasmid - Mouse IgG1 - Lucia tag

ABOUT

Heavy chain constant region expression plasmid - Mouse IgG1 - Lucia tag

pFUSE-Lucia-CHIg-mG1 is a cloning plasmid that expresses the   constant region of the murine IgG1 heavy chain. It contains a multiple cloning site upstream of the constant region to enable cloning of the heavy chain variable region.

This plasmid is   designed for antibody generation when co-transfected into mammalian cells with the light chain variable region cloned into pFUSE-CLIg.

pFUSE-Lucia-CHIg-mG1 contain  the secreted luciferase (Lucia) gene upstream of the MCS and the CH  region to serve as a tag to facilitate the detection and quantification  of recombinant antibodies.

All products are for internal research use only, and not for human or veterinary use.

SPECIFICATIONS

Specifications

Plasmid backbone
pFUSE
Purification
Ion-exchange chromatography
Reconstitution buffer
Sterile Water
Quality control

Plasmid construct has been confirmed by restriction analysis and sequencing

CONTENTS

Contents

  • Product: 
    pFUSE-Lucia-CHIg-mG1
  • Cat code: 
    pfuselc-mchg1
  • Quantity: 
    20 µg
Includes:

1 ml of Zeocin™ (100 mg/ml)

Shipping & Storage

  • Shipping method:  Room temperature
  • Storage:

    • -20°C

    Caution:

    • Avoid repeated freeze-thaw cycles

Details

pFUSE-Lucia-CHIg backbone

 

1- Schematic representation of a Lucia luciferase-tagged antibody

Lucia-tagged antibody produce with pFUSE vectors

2- Luciferase activity of Lucia-tagged anti-hTNF-α antibodies.

Luciferase activity of Lucia-tagged anti-hTNF-a antibodies

 

CHO cells were stably co-transfected with a heavy chain expressing plasmid, pFUSE-Lucia-TNF-CHIg-hA2m1 (TNF-CHIg-hA2m1) or pFUSE-Lucia-TNF-CHIg-hG1 (TNF-CHIg-hG1) and a light chain expressing plasmid, pFUSE2-TNF-CLIg-hk (TNF-CLIg-hk) to generate Lucia-tagged anti-hTNF-α antibodies, Lucia-anti-hTNF-α-hIgA2m1and Lucia-anti-hTNF-α-hIgG1, of human IgAm2 and human IgG1 isotypes, respectively. pFUSE2-CLIg-hk (CLIg-hk), which expresses no VL, and pSELECT-blasti-mcs (mcs) were used as negative controls. Supernatants
were collected and the luciferase levels determined using QUANTI-Luc™. Only the cells co-producing a Lucia-anti-hTNF-α heavy-chain and an anti-hTNF-α light chain displayed luciferase activity.

 

3- Neutralizing activity of anti-hTNF-a antibodies

Neutralizing activity of anti-hTNF-alpha antibodies

The activity of anti-hTNF-α-hIgA2m1 and Lucia-anti-hTNF-α-hIgA2m1 antibodies, purified from the supernatants of CHO transfected cells, was determined by performing an TNF-α neutralizing assay. Both antibodies display similar neutralizing activities, thus fusion of the Lucia luciferase tag at the N-terminus of the heavy chain does not alter the functionality of the antibody.

DOCUMENTS

Documents

pFUSE-Lucia-CHIg-mG1

Technical Data Sheet

Safety Data Sheet

Plasmid Sequence

Certificate of analysis

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