Analysis of protein extracts from films and gloves produced with hevea and guayule latex

Transcription

1 Analysis of protein extracts from films and produced with hevea and guayule latex M Daniau, G Olivier, A Souverain, D Pioch, S Palu, M Rossignol, S Hem, D Sauvaire, C Mourton-Gilles Biorubber for Europe in Global Perspective, Wageningen, September 25th, 212

2 Scientific context Latex allergy is due to the présence of hevea proteins in rubber goods (cleaning, nipples, toys, ) and medical devices (, catheter, syringe, ). Latex allergy is a real health problem Up to 17% of health care workers suffer from type I hevea allergy Up to 16% of intra-operative shock are due to this allergy Presence of cross allergy with some fruits (banana, kiwi, ) A solution to this problem could be to develop devices with a non allergenic latex : GUAYULE LATEX Objective Collaborative study between CIRAD and National Health product Agency To compare protein content of Guayule and Hevea device extracts

3 Determination of total protein content in films and extracts Protein quantity (µg/g) Hevea films Guayule films Powered free latex 5.35 Sterile powered free latex (CIRAD/CTTM ) Yulex (CIRAD/CTTM ) EU-PEARLS (CIRAD/CTTM ) Protein quantity in guayule films are 115 times lower than in hevea films and 9 times than in medical Gloves made with commercial guayule latex contain less protein than hevea prototype. Gloves made with CIRAD guayule latex contain large protein amounts 2

4 Hev b1 (µg/g) 1.2E E-2 Determination of allergen content in films and extracts 1.E-2 8.E-3 6.E-3 4.E-3 Allergen Hevb1 is present in the hevea films and in small amount in guayule extracts from films and 2.E-3.E+ 4.3E E-4 Hevea film Guayule film Pow ered free latex Steril pow ered free latex 6.33E E-4 Yulex EU-PEARLS 2.5E-1 Hev b3 (µg/g) 2.17E-1 2.E-1 1.5E-1 Allergen Hevb3 is present in the hevea films and in small amount in medical and guayule made with commercial latex 1.E-1 5.E-2.E+ 1.87E E-3 Hevea film Guayule film Pow ered free latex Steril pow ered free latex 7.22E E-3 Yulex EU-PEARLS 3

5 Determination of allergen content in films and extracts Hev b5 (µg/g) 5.E E-3 4.5E-3 4.E-3 3.5E-3 3.E-3 2.5E-3 2.E-3 1.5E-3 1.E E-4 Allergen Hevb5 is present in the hevea 5.E-4.E+ 2.33E-5 Hevea film Guayule film Pow ered free latex 6.71E-5 Steril powered free latex Yulex EU- PEARLS 1.2E-2 Hev b6.2 (µg/g) 1.2E-2 1.E-2 8.E-3 6.E-3 5.5E-3 Allergen Hevb6.2 is present in the hevea films and but also in guayule films and 4.E-3 2.E E-4 2.7E-3 2.1E E E-3.E+ Hevea film Guayule film Pow ered free latex Steril powered free latex Yulex EU- PEARLS 4

6 Identification of proteins in guayule films extracts Using WB and labelling with anti-hevea Mabs (Hevb1,Hevb3,Hevb5 and Hevb6.2) Hevb1 and Hevb6.2 were detected in guayule films extracts Using LC ESI MS/MS mass spectrometry: collaboration with INRA Some hevea allergens were identified Rubber elongation factor Hevb1 Glucan endo1,3 beta glucosidase: Hev b2 Hevamin A : Hev b14 Pro-Hevein: Hev b6.1 One guayule protein was identified Allene oxyde synthase 5

7 Conclusion Protein quantity in guayule films is about 1 times lower than in hevea films Only Hevb1 and Hevb6.2 were detected in guayule films using Fit Kits Gloves made with commercial guayule latex contain less protein than hevea But, made with CIRAD guayule latex contain large protein amounts Small amounts of Hevb1, Hevb3 and Hevb6.2 were detected in guayule glove prototypes Using mass spectrometry, four hevea allergens were found in guayule film extract Hevb1, Hevb2, Hevb14 and Hev b6.1 We detected Hevb6.2 (hevein) by ELISA or Hevb6.1 (pro-hevein) by MS Identification of a guayule specific protein (Parthenium argentatum) : Allene oxyde synthase 6

8 Thank you for your attention 7

9 Avertissement Lien d intérêt : personnel salarié de l ANSM (opérateur de l Etat). La présente intervention s inscrit dans un strict respect d indépendance et d impartialité de l ANSM vis-à-vis des autres intervenants. Toute utilisation du matériel présenté, doit être soumise à l'approbation préalable de l ANSM. Warning Link of interest: employee of ANSM (State operator). This speech is made under strict compliance with the independence and impartiality of ANSM as regards other speakers. Any further use of this material must be submitted to ANSM prior approval.