Pollen grain column chromatography: quantitation and biochemical analysis of ragweed-pollen solutes

J Allergy Clin Immunol. 1988 Jun;81(6):1126-34. doi: 10.1016/0091-6749(88)90880-9.


The kinetics, quantitative yield, and sequence of solute release during the extraction of allergenic substances from short ragweed (Ambrosia artemisiifolia) pollen were compared with a conventional batch-type method and the novel technique of pollen grain column chromatography. With the batch method, 14.6 +/- 1.7 mg of pollen solutes were eluted per 100 mg of dried defatted pollen in 1 minute; the 24-hour solute yield was 27.4 +/- 2.7 mg. With the column method, 3.7 +/- 1.3 mg of pollen solutes were eluted in 1 minute; the 24-hour solute yield was 29.3 +/- 2.1 mg. The kinetics of solute release with the column method were modeled as the simultaneous first-order elution of ragweed-pollen solutes into three hypothetical compartments. The theoretical initial solute concentration was 50 gm/L. The isoelectric focusing patterns, optical properties, distributions of enzymes, Ra5, and antigen E activities were consistent with the sequential separation of ragweed-pollen solutes and the three compartment model. Enzyme activities were eluted either maximally in the first minute (phosphatases and N-acetyl-beta-glucaminidase) or delayed until 10 minutes (leucine aminopeptidase). Ra5 was eluted rapidly, whereas antigen E was eluted during a more prolonged period. Pollen grain chromatography provides a simple, reproducible method for studying pollen solute release.

MeSH terms

  • Acid Phosphatase / metabolism
  • Alkaline Phosphatase / metabolism
  • Allergens*
  • Antigens, Plant
  • Cell Extracts / analysis
  • Chromatography / methods*
  • Esterases / metabolism
  • Isoelectric Focusing
  • Kinetics
  • Phosphoric Monoester Hydrolases / metabolism
  • Plant Proteins*
  • Pollen / metabolism
  • Substrate Specificity


  • Allergens
  • Amb a I protein, Ambrosia artemisiifolia
  • Antigens, Plant
  • Cell Extracts
  • Plant Proteins
  • group 5 allergens, Ambrosia
  • Esterases
  • Alkaline Phosphatase
  • Acid Phosphatase
  • Phosphoric Monoester Hydrolases