Method for preparing type b Haemophilus influenzae and polysaccharide conjugate vaccine

Abstract

The invention discloses a method for preparing a type b Haemophilus influenzae and polysaccharide conjugate vaccine. The method comprises the following steps: S1, performing gel filtration chromatography purification of polysaccharide to obtain macromolecular Hib capsular polysaccharide; and S2, preparation of a polysaccharide-TT conjugate, which comprises the following sub-steps: S21, adding CNBr into a macromolecular Hib capsular polysaccharide liquor to activate for 15-25 minutes, then, adding adipic dihydrazide to derivate for 10-20 minutes to remove residual CNBr so as to obtain a polysaccharide-AH derivative, and S22, adding TT liquor to the polysaccharide-AH derivative, uniformly mixing and placing in an ice-bath, and adding EDAC powder, and separating, purifying, degerming and filtering the reactant to obtain the finished product. The method disclosed by the invention has the beneficial effects that the yield of qualified conjugates is improved, the raw material consumption in the conjugation process is reduced, and the stability of the production process and the consistency of quality of finished products are improved.

Description

technical field [0001] The invention relates to the technical field of vaccine preparation methods, in particular to a method for preparing Haemophilus influenzae type b polysaccharide-conjugated vaccine. Background technique [0002] Haemophilus influenzae (Haemophilus Influenza type b, Hib) is still the main pathogen causing human invasive diseases so far, most of which are caused by Haemophilus influenzae type b (Hib). Hib meningitis ranks first among bacterial meningitis, and 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 under the age of 5 , especially under the age of 2. It is estimated that it causes at least 3 million cases of severe disease each year, of which about 386,000 people die, and has become a major global public health problem. In developing countries, Hib pneumonia plays an impo...

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Problems solved by technology

[0006] However, because there are a large amount of small molecule polysaccharides with a molecular weight lower than 700kd in the refined Hib capsular polysaccharide prepared, the molecular weight of this part of the polysaccharide is small after being combined with the carrier protein, and it is separated after gel filtration chromatography, resulting in a high concentration during the binding process. The yield of molecular polysaccharide-protein conjugates is very low, and the yield of Hib capsular polysaccharide polysaccharide in this preparation method in the industry is only 4-6%.

In addition, due to the uneven molecular size distribution of the refined Hib capsular polysaccharide obtained in the preparation process, there is a large difference in the molecular size distribution of each batch, resulting in different physical and chemical indicators of each batch of polysaccharide-protein conjugated vaccine stock solution, which affects the quality of the production process. Stability, batch-to-batch stability of finished vaccine and consistency of finished vaccine quality

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