Isolation of monoclonal immunoglobulin from plasma of patients with multople myeloma using affinity chromatography
| Authors | |
|---|---|
| Year of publication | 2002 |
| Type | Article in Proceedings |
| Conference | Zborník XVII.Biochemický zjazd |
| MU Faculty or unit | |
| Citation | |
| Field | Oncology and hematology |
| Keywords | multiple myeloma; IgG; affinity chromatography; G-protein |
| Description | Multiple myeloma is characterized by the proliferation of a malignant plasma cell clone and accounts for approximately 10% of hematological malignancies. High-dose chemotherapy with autologous transplantation has improved the survival of patients with myeloma, hovewer, relapse is inevitable (1,2). Recent research has targeted the minimal residual disease. One of the therapeutical options is this setting is immunotherapy using specific tumor antigen as a vaccine. The idiotype of myeloma immunoglobulin (Id protein) is specifically expressed by malignant cells and can be used as a tumor antigen with the aim to induce cytotoxic T-lymphocyte response against myeloma cells (3,4,5,6). For purification of Id protein from plasma of relapsed IgG myeloma patients who were before reinduction treatment, plasma was precipitated with a saturated solution of ammonium sulfate. After precipitation and dialysis in saline at pH 7.3 to remove ammonium sulfate, the Id protein was separated on protein G immobilized on agarose. To breach the bound between the Id-protein and the protein G, 0.2M glycine (pH 2.7) was used and resulting fractions were immediately neutralized in 1M Tris-base. SDS-polyacrylamide gel electrophoresis was used to determine the purity of Id protein. The isolation was carried out in 10 patients. Using columns with a capacity of 30 mg/ml protein G and the flow rate of 1-2 drops per 10 seconds, 47% of the plasma content of Id protein was extracted (range: 22 to 89%). The initial concentration of monoclonal immunoglobulin in plasma did not influence the yield of the separation. The amount of Id protein extracted in each isolation was sufficient to a prepare Id-KLH conjugate that was used as the specific antigen in preclinical experiments. |
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