Original article
Pollen grain column chromatography: Quantisation and biochemical analysis of ragweed-pollen solutes

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Abstract

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-β-glucaminidase) or delayed until 10 mintues (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

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  • Cited by (0)

    1

    From the Departments of Medicine and Microbiology-Immunology, Duke University, Durham, N.C.

    From the Greer Laboratories, Lenoir, N.C.

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