Download Electromanipulation in Hybridoma Technology: A Laboratory by Carl A.K. Borrebaeck, Inger Hagen PDF

By Carl A.K. Borrebaeck, Inger Hagen

This quantity offers a cutting-edge review of the options of electrofusion and electroporation utilized in the construction of either human and mouse monoclonal antibodies. it's the purely paintings to be had to aspect the pitfalls of electrofusion and to supply professional assistance in overcoming them. With its exact protocols, this "how-to" handbook enables the winning construction of achievable telephone hybrids and transfectants.

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However, because no significant effects on membrane transport properties and cell viability are observed, it must be assumed that these changes in membrane structure are minor. Such small changes in membrane structure can be envisaged if the membrane and/or cytoskeletal proteins are (partly) dissolved by the osmotic stress, resulting in a higher mobility of membrane components and probably in the emergence of protein-free lipid domains. The experiments reported here force the conclusion that disruption of restraining proteins by osmotic means is more advantageous than by field pulses alone.

C. Human Transfectants/Monoclonal Antibody Production We have generated a panel of human transfectants by stimulating mixed population of lymphocytes with mitogens and transfecting these stimulated/activated cells with oncogenic DNA/oncogene(s). The transfectants displayed various phenotypes, indicating that they were immortalized at different stages of their differentiation cascade. The karyotype of these cells appeared normal. The observations that these transfectants are also phenotypically stable and secrete immunoglobulin make them good candidates for routine monoclonal antibody production.

1989. Efficient generation of stable antibody forming hybridoma cells by electrofusion. Hybridoma 8:107-115. 33. , and Zimmermann, U. 1979. Particles in a homogeneous electrical field: A model for the electrical breakdown of living cells in a Coulter Counter. Bioelectrochem. Bioenerg. 6:349-384. 34. , Schnettler, R, and Zimmermann, U. 1987. Parameters controlling yeast hybrid yield in electrofusion: The relevance of pre-incubation and skewness of the size distributions of both fusion partners. Biochim.

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