Optimization and extraction process of whole length polyclonal rabbit anti-human SMN antibody

Abstract

The present invention discloses the optimization and extraction process of whole length polyclonal rabbit anti-human SMN antibody, and belongs to the fields of early genic diagnosis of spinal muscular atrophy, SMN protein function research and disease prognosis judgment. The whole length polyclonal rabbit anti-human SMN antibody is prepared with the whole length SMN protein of 1-294 amino acids, and points to all functional regions of SMN protein. The preparation process includes preparation of whole length code region cDNA segment of health human, primer design and His gene fusion vector selection, preparation of the target gene segment, purification of His gene fusion protein and collection of target protein, and purification. The antibody of the present invention has certain specificity and sensitivity, and may play positive part in said fields.

Description

technical field [0001] The invention relates to methods for early gene diagnosis of diseases, SMN protein function research and disease prognosis judgment, in particular to the optimization and extraction of rabbit anti-human SMN full-length polyclonal antibodies for spinal muscular atrophy. Background technique [0002] Spinal muscular atrophy (SMA) is a hereditary disease characterized by degeneration of α-motor neurons in the anterior horn of the spinal cord, weakness and atrophy of the proximal limbs. Weakness and muscle atrophy, the proximal end is heavier than the distal end, and the lower extremity is heavier than the upper extremity. SMA with onset in childhood is autosomal recessive, with a population incidence of 1 / 6000-1 / 10000 and a carrier frequency of 1 / 40-1 / 60. According to the age of onset and the severity of the disease, SMA with onset in childhood is clinically divided into 3 types: children with type I usually die of respiratory muscle paralysis within 2 y...

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Problems solved by technology

[0005] Retrieval of relevant materials including Chinese patents shows that there is no ready-made SMN antibody in China at present, while the SMN antibodies prepared by foreign scholars are all carboxy-terminal or amino-terminal polypeptide antibodies, and only dozens of amino acids need to be synthesized for the preparation of such antibodies The polymorphic polymorphic deimmunized rabbits, the preparation process is relatively simple and fast, but because it cannot cover the three different key regions of the SMN protein at the same time, the sensitivity and specificity of this type of antibody are low, which limits the depth of antibody use And breadth, such as immunofluorescent staining of cells only or immunohistochemistry only, and prone to false positive or false negative results, so not the best SMN antibody

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