Protein extraction from cypress pollen with Minilys - Bertin Instruments

    Environment

    Protein extraction from cypress pollen with Minilys

    Sources: ESPCI, LSABM, Allergy & Environment, France

    Context

    The common cypress pollen (Cupressus sempervirens) is becoming an increasing cause of respiratory allergy in Europe and some regions worldwide. Because of its particular structural features and physico-chemical composition, cypress pollen is one of the most pollen difficult to analyze in terms of protein content and therefore allergens. In aqueous media, the external wall (exine) cracks in a few minutes under the effect of swelling of the intine (inner wall) particularly rich in polysaccharide. Few proteins are then extracted in aqueous conditions. The dry milling may be a good alternative for the extraction of cypress pollen proteins and to generate fragments of smaller sizes for experiments and ultrastructural analysis of immuno-reactivity [1].

    [1] Y. Shahali, J.-P. Sutra, G. Peltre, D. Charpin, H. Sénéchal,P. Poncet. IgE reactivity to Common cypress (C. sempervirens)pollen extracts: evidence for novel allergens. WAO Journal.2010, 3, 229-234.

    Results

    Microscopic observation (Figure 1) confirms that the Cupressus pollen dry milling with CK14 beads during 3x30sec seems suited to generate smalls fragments of pollen. The downstream analysis (results not shown) show that Minilys dry milling is very efficient to extract quickly high concentration of proteins. However, the protein extracted by dry milling were different from the protein extracted by overnight incubation in aqueous media (PBS). The dry milling is recommended in respect of cypress pollen due to his feature gelling in aqueous media. Other pollen grains such as birch or grass pollen can be grind in liquid media.

     

    Microscopic observation (X40). Whole Cupressus pollen grain before grinding, dry observation (1a). Cupressus pollen grain after 1x30s dry grinding, observation in water (1b). Cupressus pollen grain after 3x30s dry grinding, observation in water (1c).

    Microscopic observation (X40). Whole Cupressus pollen grain before grinding, dry observation (1a). Cupressus pollen grain after 1x30s dry grinding, observation in water (1b). Cupressus pollen grain after 3x30s dry grinding, observation in water (1c).

     

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