High-density culture and production method of bacterium capsular polysaccharide with haemophilus influenzae type b

Abstract

The invention discloses a high-density culture and production method of bacterium capsular polysaccharide with haemophilus influenzae type b. The method comprises following steps: subculturing Hib strains from a first generation to a fourth generation, carrying out culture in a seeding tank, carrying out fermentation and culture in a fermentation tank, killing bacteria, acquiring a supernatant, and purifying the supernatant. Beneficial effects of the invention are as follows: (1) by culturing Hib bacteria to a high concentration, output of Hib capsular polysaccharide is improved by 50% to 100% on the basis of the output of Hib capsular polysaccharide produced by the traditional way, and produced capsular polysaccharide meets the national standard; (2) by controlling nucleic acids, protein, and other impurities in a zymotic fluid to a low level, the number of times of phenol extracting for removing impurity protein is reduced for relatively low protein content, thereby reducing phenol amount used in subsequent purifying processes, reducing environmental pollution, and reducing production cost; and (3) output of polysaccharide is stable, and content of refined glycoprotein, endotoxin, and other impurities after the purifying is substantially lower than the national control standard.

Description

technical field [0001] The invention relates to a method for high-density culture of Haemophilus influenzae type b to produce bacterial capsular polysaccharide. Background technique [0002] Today, Haemophilus influenzae (Hi) is still the main pathogenic bacteria causing human invasive diseases, most of which are caused by Haemophilus influenzae type b (Hib). The harm of Hib meningitis ranks first among bacterial meningitis, and it has the characteristics of high morbidity, high mortality and high disability rate; another characteristic is that the age of onset is young, the infection rate of infants is high, and the age of onset is mainly concentrated in the age of 5 under the age of 2, especially under the age of 2. According to the World Health Organization (WHO) report in 2006, Hib meningitis causes at least 3 million cases of severe disease every year worldwide, of which about 386,000 people die, which has become a major global public health problem. In developing cou...

AI Technical Summary

Problems solved by technology

The yield of Hib capsular polysaccharide in the traditional production method is low, and the cost in the vaccine production process is high, which is one of the reasons for the high price of Hib vaccine

At the same time, since the Hib fermentation broth contains more impurities such as nucleic acids and proteins, the difficulty of subsequent purification is increased. In order to obtain high-purity Hib capsular polysaccharides, the purification process can only be increased, which further reduces the concentration of Hib capsular polysaccharides. The yield, so that the yield further decreased

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