Natural Der p 1 Molecular Reference Std. (MRS-NDP1-1)

The Natural Der p 1 MRS is intended to serve as reference standard to determine the Der p 1 content of allergen preparations from house dust mite (Dermatophagoides pteronyssinus) by immunoassay.

Allergen: Natural Der p 1 (Dermatophagoides pteronyssinus allergen 1)

Source: D. pteronyssinus culture

Mol. Wt: 24 kD

Purification: From spent mite culture by multi-step affinity chromatography.

Composition: 10µg natural Der p 1, determined by Amino Acid Analysis, freeze dried in sealed glass vial.

SDS-PAGE: See inset. Silver-stained SDS-PAGE under non-reducing conditions shows a single band at 24 kD.

ELISA: Immunoreactive in Der p 1 specific ELISA. No trace contamination with Der p 2 was detected by ELISA.

Purity: >95% purity by in-solution LC-MS/MS after tryptic digest.

Isoforms: Analyzed by LC-MS/MS. Main Isoforms: Der p 2.0110 (43%) and Der p 2.0101 (37%). Other Isoforms present (20%): Der p 2.0103, Der p 2.0104, Der p 2.0114.

Formulation: Prior to lyophilization, natural Der p 1 was adjusted to 50 mM volatile ammonium bicarbonate with 3% trehalose.

Storage: Store at -20ºC

Product Resources: Natural Der p 1 Molecular Reference Standard Certificate of Analysis

Allergens are provided for research and commercial use in vitro: not for human in vivo or therapeutic use.

Reconstitution:

• Allow vial to reach room temperature before use
• Tap vial gently to collect all material at the bottom
• Using a sterile syringe reconstitute the MRS to desired concentration by injecting a suitable volume of a buffer of choice (e.g. PBS, pH 7.4 or 1% BSA/50% glycerol/PBS, pH 7.4).
• Mix by gently swirling the vial until content is completely dissolved.
• Adding 1ml of buffer will result in a Der p 1 concentration of 10,000ng/ml.

References:

1. Gough L, Schultz O, Sewell HF, Shakib F. The cysteine protease activity of the major house dust mite allergen Der p 1 selectively enhances the immunoglobulin E antibody responses. J Exp Med 1999; 190:1897-1901.
2. Wan H, Winton HL, Soeller C, Tovey ER, Gruenert DC, Thompson PJ, Stewart GA, Taylor GW, Garrod DR, Cannell MD, Robinson C. Der p 1 facilitates transepithelial allergen delivery by disruption of tight junctions. J Clin Invest 1999; 104:123-133.
3. Chruszcz M, Chapman MD, Vailes LD, Stura EA, Saint-Remy JM, Minor W, Pomés A. Crystal structures of mite allergens Der f 1 and Der p 1 reveal differences in surface- exposed residues that may influence antibody binding. J Mol Biol. 2009; 386:520-530.
4. Chapman MD et al. The European Union CREATE project: a model for international standardization of allergy diagnostics and vaccines. J Allergy Clin Immunol. 2008;122:882-889.
5. van Ree R et al. The CREATE project: development of certified reference materials for allergenic products and validation of methods for their quantification. Allergy. 2008;63(3):310-26.
6. Kaul S et al. Regulatory environment for allergen-specific immunotherapy. Allergy 2011;66:753-64.
7. Chapman MD and Briza P. Molecular approaches to allergen standardization. Curr Allergy Asthma Rep. 2012;12:478-84.