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The Hoffmann-La Roche Inc. patent solves the following problem:
Since the establishment of the hybridoma technology (Cole, SPC, et al, Monoclonal antibody and Cancer Therapy, Alan R. Liss, p 77 (1985); …. and Boerner, P., et al, J. Immunol 147 (1991 ) 86-95), monoclonal immunoglobulin appeared to play an important role in scientific research, human healthcare and diagnostics. Therefore, the generation of monoclonal, especially therapeutic, immunoglobulin a field undergoing intensive research. In its honor, the hybridoma technology and phage display technology (Hoogenboom, HR, and Winter, G., J. mol Biol 227 (1992) 381-388; Marks, JD, et al, J. mol Biol … 222 (1991) 581-597) was, among others, two commonly used technology for the generation of monoclonal immunoglobulin. Hybridoma technology have the strong clones is a hurdle, thus, diminishing differences in antibody, as only a limited number of B-cells successfully fused, propagated and afterward described. Similarly, one drawback of phage or yeast display-based combinatorial library methods are randomly paired immunoglobulin heavy and light chains. The dissociation of the original heavy and light chain pairing, and non-cognate pairing, requires the screening of a large number of immunoglobulin cells to recognize the heavy and light chains paris high aunt. Moreover, the non-cognate pairs may show no cross-reactivity to human antigens. In the end, the genetic diversity of target-specific immunoglobulin identified in the selection and screening of combinatorial libraries are often limited because of inherent selection bias.
Our analysis of this patent is as follows:
Hoffmann-La Roche Inc.’s patent US 9399670 B2 deals with Method for obtaining immunoglobulin encoding nucleic acid.
The present invention is directed to a method for obtaining a nucleic acid encoding an immunoglobulin variable domain from a cell comprising the following steps: performing a first polymerase reaction in three to six 5-primer and a 3-primer, performing products in the polymerase chain reaction with a second polymerase reaction thirteen to sixteen 5-primer a 3-primer, where the distance from the binding sites in primer used in the second polymerase chain reaction is reduced compared to the first chain reaction polymerase.
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