Hybridoma technology

The myeloma cell line that is used in this process is selected for its ability to grow in tissue culture and for an absence of antibody synthesis.

They shared the Nobel Prize of 1984 for Medicine and Physiology with Niels Kaj Jerne, who made other contributions to immunology.

The myeloma cells are selected beforehand to ensure they are not secreting antibody themselves and that they lack the hypoxanthine-guanine phosphoribosyltransferase (HGPRT) gene, making them sensitive (or vulnerable) to the HAT medium (see below).

Hence, unfused myeloma cells die, as they cannot produce nucleotides by the de novo or salvage pathways because they lack HGPRT.

Unlike western blot assays, immunoprecipitation-mass spectrometry facilitates screening and ranking of clones which bind to the native (non-denaturated) forms of antigen proteins.

Once a hybridoma colony is established, it will continually grow in culture medium like RPMI-1640 (with antibiotics and fetal bovine serum) and produce antibodies.

[2] Multiwell plates are used initially to grow the hybridomas, and after selection, are changed to larger tissue culture flasks.

[2] By using culture supernatant or a purified immunoglobulin preparation, further analysis of a potential monoclonal antibody producing hybridoma can be made in terms of reactivity, specificity, and cross-reactivity.

[2] Monoclonal antibodies are especially useful in distinguishing morphologically similar lesions, like pleural and peritoneal mesothelioma, adenocarcinoma, and in the determination of the organ or tissue origin of undifferentiated metastases.

[2] One study[5] performed a sensitive immuno-histochemical assay on bone marrow aspirates of 20 patients with localized prostate cancer.

Although immuno-cytochemistry using tumor-associated monoclonal antibodies has led to an improved ability to detect occult breast cancer cells in bone marrow aspirates and peripheral blood, further development of this method is necessary before it can be used routinely.

[6] One major drawback of immuno-cytochemistry is that only tumor-associated and not tumor-specific monoclonal antibodies are used, and as a result, some cross-reaction with normal cells can occur.

They concluded from the results that immuno-cytochemical staining of bone marrow and peripheral blood is a sensitive and simple way to detect and quantify breast cancer cells.

The use of monoclonal antibodies (mAbs) specific for cytokeratins can identify disseminated individual epithelial tumor cells in the bone marrow.

The twelve control aspirates from patients with benign prostatic hyperplasia showed negative staining, which further supports the specificity of CK18 in detecting epithelial tumour cells in bone marrow.

[11] One study has reported the isolation of a hybridoma cell line (clone 1E10), which produces a monoclonal antibody (IgM, k isotype).

Monoclonal antibodies (mAbs) specific for cytokeratins can detect disseminated individual epithelial tumour cells in the bone marrow.