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Establishment a CHO Cell Line Expressing Human CD52 Molecule.

Kadijeh Tati¹, Mahsa Yazdanpanah-Samani², Amin Ramezani³

¹Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical sciences, Shiraz, Iran; Department of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.

Reports of Biochemistry & Molecular Biology

|January 11, 2017

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This summary is machine-generated.

Researchers cloned and expressed the human CD52 gene in CHO cells, creating a CD52-positive cell line. This advancement aids in developing diagnostic tools and potential antibody therapies for CD52-related conditions.

Area of Science:

Molecular Biology
Immunology
Cell Biology

Background:

CD52 is a GPI-anchored glycoprotein found on lymphocytes and monocytes.
Soluble CD52 in chronic lymphocytic leukemia (CLL) plasma may serve as a tumor marker.
CD52's precise function is unknown but may influence T-cell migration and activation.

Purpose of the Study:

To clone and express the human CD52 gene in a Chinese Hamster Ovary (CHO) cell line.
To investigate the functional role of CD52 in more detail.

Main Methods:

CD52 gene amplification from Raji cell line cDNA using specific primers.
Cloning the amplified gene into the pBudCE4.1 vector, creating the pBudKT1 construct.
Transfection of CHO-K1 cells with the construct via electroporation and confirmation of expression using RT-PCR and flow cytometry.

Keywords:

CD52 Recombinant DNA Therapeutic and diagnostic proteins

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Main Results:

Successful amplification and cloning of the CD52 gene, confirmed by PCR and sequence analysis.
Detection of CD52 mRNA in transfected CHO cells via RT-PCR.
Flow cytometry confirmed CD52 protein expression on the surface of 78.4% of transfected cells.

Conclusions:

A human CD52-expressing CHO cell line (CHO-CD52) was successfully established.
This cell line serves as a foundation for producing therapeutic monoclonal antibodies against CD52.
The development facilitates detection systems for soluble CD52 in biological fluids.