Karakterizacija alergenov v mesnih izdelkih

Meat allergy is a less common and poorly studied type of food allergy. Nevertheless, its prevalence is not negligible. An important factor that can affect meat allergenicity is its processing into meat products. In the first part of this master thesis, we identified putative allergens (serum albumin, myosin light chain, parvalbumin) in various extracts from pork meat products and pork meat by means of immunoblotting and the use of antibodies specific for these proteins. Allergen identity was confirmed by mass spectrometry. In the second part, we produced myosin light chain isoforms 1 and 3 and parvalbumin in bacterial expression systems. We evaluated their binding capacity of IgE antibodies from patients’ sera, their thermal stability, folding, aggregation, and resistance to digestion by enzymes of the gastrointestinal tract. Characterisation of recombinant allergens confirmed their comparability with naturally occurring proteins and thus adequacy for their use in further studies. The identified allergens cannot be regarded as major pig allergens, but their thermal stability and partial resistance to digestion imply that preparation and processing of meat do not affect their allergenicity and can thus these allergens consequently be retained in pork meat products as well.Alergija na meso je manj pogost in slabše raziskan tip alergije na hrano, a njena razširjenost kljub temu ni zanemarljiva. Pomemben dejavnik, ki lahko vpliva na alergenost mesa, je predelava le-tega v mesne izdelke. V prvem delu te magistrske naloge smo identificirali domnevne alergene (serumski albumin, miozinsko lahko verigo, parvalbumin) v ekstraktih različnih svinjskih mesnih izdelkov in svinjskega mesa z metodo prenosa po Westernu in uporabo za te proteine specifičnih protiteles. Njihovo identiteto smo potrdili z metodo masne spektrometrije. V drugem delu smo izoformi 1 in 3 miozinske lahke verige ter parvalbumin proizvedli v bakterijskem ekspresijskem sistemu. Preverili smo njihovo sposobnost vezave protiteles IgE iz seruma alergikov, njihovo toplotno stabilnost, zvitje, potencialno tvorbo višjih molekularnih struktur ter odpornost na razgradnjo z encimi prebavnega trakta. Karakterizacija rekombinantnih alergenov je potrdila njihovo podobnost z naravno prisotnimi proteini in tako primernost za uporabo v nadaljnjih raziskavah. Identificiranih alergenov ne moremo uvrstiti med ključne prašičje alergene, a potrjena stabilnost pri povišani temperaturi in deloma v prisotnosti prebavnih encimov nakazujeta, da postopki priprave in predelave mesa njihove alergenosti ne zmanjšajo in se potencialni mesni alergeni lahko ohranijo tudi v svinjskih mesnih izdelkih