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[Purification and analysis of the immunoactivity of humulus pollen allergen]. | LitMetric

[Purification and analysis of the immunoactivity of humulus pollen allergen].

Sichuan Da Xue Xue Bao Yi Xue Ban

Respiratory Departments, Second Affiliated Hospital of Medical College, Xi'an Jiaotong University, Xi'an 710004, China.

Published: November 2008

AI Article Synopsis

  • The study aimed to purify humulus pollen allergen and analyze its allergenic and immunogenic properties.
  • Crude pollen extracts were separated using gel filtration methods, resulting in various protein fractions, with a focus on those containing significant allergenic proteins.
  • Results showed that different protein fractions had varying molecular weights and inhibition rates, with certain fractions being more reactive to specific immunoglobulins (sIgG and sIgE) in patient serum, indicating differing levels of allergenicity.

Article Abstract

Objective: To purify the humulus pollen allergen and study the allergenicity and immunogenicity of it.

Methods: Crude humulus pollen extracts were purified by gel filtration with Sephadex G-75 and Sephacryl S-200HR. Various fractions of the allergen protein were collected respectively. The molecular weights of protein were confirmed by SDS-PAGE. The inhibition rate and reaction rate with sIgG and sIgE of patient's serum were determined by ELISA inhibition test and western blotting.

Results: Two peaks were obtained from crude humulus pollen extracts by gel filtration of Sephadex G-75. The first peak contained most of protein while the second peak contained little protein and lots of pigments. So the second peak was thrown away. P solution which contained the first peak and valley fraction were purified by gel filtration of Sephacryl S-200HR and four components that included the 1st peak, the valley, the 2nd peak and the end fraction were obtained. The results of electrophoresis demonstrated that purified humulus pollen contained more than 20 kinds of protein with the molecular weights ranged from 5.0 x 10(3) to 97.4 x 10(3). The fraction of the 1st peak contained protein with the molecular weights ranged from 43 x 10(3) to 97.4 x 10(3), and the fraction of the valley and the 2nd peak contained protein with the molecular weights ranged from 5.0 x 10(3) to 43 x 10(3). The fraction of the end contained protein with the molecular weights lower than 5.0 x 10(3). The results of ELISA inhibition test showed that the inhibition rate of the 1st peak, the valley, the 2nd peak and the end fraction to sIgG were 68%, 70%, 95%, 5% respectively, and those to sIgE were 25%, 64%, 71%, 11% respectively. The results of western blotting demonstrated that the reaction rate of the 1st peak, the valley, the 2nd peak and the end fraction with sIgG of patients' serum were 65.63%, 78.13%, 87.50%, 6.25% respectively, and those with patients' sIgE were 25.00%, 71. 88%, 84.38%, 15.63% respectively.

Conclusion: Humulus pollen contained more than 20 kinds of protein. The proteins with molecular weights ranged from 5.0 x 10(3) to 43 x 10(3) were the major allergen with strong allergenicity and immunogenicity. The proteins with molecular weights ranged from 43 x 10(3) to 97.4 x 10(3) were subordinated allergen with strong immunogenicity and weak allergenicity.

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