[Construction and expression of chimeric anti-human CD20 monoclonal antibody].

Aim: To construct the eukaryotic expression vector of chimeric anti-human CD20 monoclonal antibody (mAb) and realize its expression.

Methods: The light- and heavy-chain genes were amplified from hybridoma cell line 1-28 secreting anti-human CD20 mAb by RT-PCR and were cloned to T vector and sequenced. Proteins of mAb 1-28 were separated by reducing SDS-PAGE.

[Cloning and expression of soluble B lymphocyte stimulator].

Aim: To clone and express soluble B lymphocyte stimulator (sBLyS).

Methods: Total RNA was isolated from peripheral blood mononuclear cells, and used to synthesize cDNA by reverse transcription. sBLyS cDNA was amplified by PCR with specific primers and inserted into a prokaryotic expression vector pET-30a.

A novel neutralizing monoclonal antibody against cell-binding polypeptide of ricin.

One strain of neutralizing monoclonal antibody (MAb) against cell-binding polypeptide of ricin, named 3E1, was generated efficiently. The antibody recognized the linearity epitope of RTB located in a toxin structure domain characterized by Western blotting. The safe period of mice for intraperitoneal injection of 100 microg of antibody was 20 min after intraperitoneal injection of 2 microg of Ricin (10 times LD50).

[Experimental study of adoptive immunotherapy using CD3AK cells].

Aim: To analyze the immunological properties and biological activity of a monoclonal antibody (mAb) against CD3 molecule(yCD3), and to observe the tumor-suppressive activity of CD3AK cells in vitro and in vivo.

Methods: FCM was used to test the specificity of yCD3 and the immunological phenotype and cytokine production of CD3AK. 3H-TdR assay was used to measure the transformation of lymphocytes activated by yCD3.

[Expression and Characterization of Single-chain Fv-Fc Fusion Protein against Human P185(erbB2)].

In order to increase therapeutic effects and decrease immunogenicity of mouse McAb, the single-chain Fv (scFv) created by fusing the light and heavy chain variable region genes of anti-human P185(erbB2) McAb was conjugated to the Fc gene of human IgG1 to construct a scFv-Fc fusion gene. The scFv-Fc fusion gene was cloned into the expression vector pCIDN. The scFv-Fc fusion protein was synthesized as secreted two-chain molecule in CHO cells, and purified by affinity chromatography on recombinant protein A.

IL-6 inhibits apoptosis of human myeloma cell line XG-7 through activation of JAK/STAT pathway and up-regulation of Mcl-1.

Background & Objective: IL-6 can protect myeloma cells from apoptosis induced by various stimuli. A series of intracellular molecules might participate in this process. Our aim in this study is to investigate which anti-apoptotic Bcl-2 family proteins (Bcl-2, Bcl-XL, Mcl-1) and which signal transduction pathway(JAK/STAT, Ras/MAPK, PI-3K/Akt) can mediate the anti-apoptotic effect of IL-6 on a human myeloma cell line XG-7.

Cloning and Expression of Human CD20 Gene on NIH-3T3 Cell Membrane.

CD20 is a specific antigen expressed on normal and neoplastic B cells exclusively. Recent researches showed that in B cell leukemia, CD20 was over-expressed. Therefore monoclonal antibody (McAb) to CD20 may be of clinical value in diagnosis and treatment of some leukemias and lymphomas.