🍎 Detection and quantification of food allergens and allergenic ingredients in foodstuffs by antibody-based methods (ELISA and other immunoassays) — framework for qualitative and quantitative methods, guidelines and performance criteria for methods detecting proteins or peptides that are markers of the presence of allergy-provoking foods, requirements for laboratories, procedure, interpretation of results and factors influencing the result; used together with EN 15842; PN-EN 15633-1:2020-03

Food chemistry PN-EN 15633-1

Overview

WHAT THE STANDARD COVERS. We read the text of the standard in the iTeh sample of SIST EN 15633-1:2019 (Slovenian adoption of EN 15633-1:2019 of October 2019, English text) — PN-EN 15633-1:2020-03 adopts EN 15633-1:2019 as identical (PKN search): contents, European foreword, introduction, Clauses 1–2 and Clause 3 up to 3.3.2 — the sample ends on page 7. We do not read the rest of the definitions, general requirements for laboratories (4), procedure (5), interpretation and expression of results (6), parameters influencing results (7) or the test report (8); we know the titles from the contents.

ACCORDING TO THE TEXT OF EN 15633-1:2019. The standard was prepared by CEN/TC 275 “Food analysis — Horizontal methods” (secretariat DIN); CEN approved it on 12.08.2019, national adoption and withdrawal of conflicting standards by April 2020 at the latest. It supersedes EN 15633-1:2009; significant changes: updated terms and definitions (3), pre-requisite requirements for analysis (5), method validation parameters (6) and specific influences on results (7). Introduction: a specific protein, group of proteins or peptides derived from them can serve as a marker for the presence of foods or ingredients provoking allergic reactions; the document describes qualitative detection and/or quantification of such analytes in the protein extracted from the sample; appropriate extraction procedures are part of each method; the focus is on antibody-based methods; “shall” — requirement, “should” — recommendation, “may” — permission, “can” — possibility. Scope (1): framework for qualitative and quantitative methods for determining food allergens and allergenic ingredients in foods using antibody-based methods; general guidelines and performance criteria for such methods; other methods can also detect these proteins; the guidelines, minimum requirements and criteria are to ensure reproducible results by different analysts in private and official laboratories and in on-site testing; the document is used in addition to EN 15842; it could also apply to other sample types. Reference (2): EN 15842 (detection of food allergens — general considerations and validation of methods). Definitions (3): denaturation of proteins — treatment (thermal, chemical, enzymatic or other) changing secondary, tertiary and quaternary structure so that only the primary structure or parts of it remain; it can modify antigenic properties; cross-linkage of proteins — chemical reaction or physical interaction between proteins, can modify extractability of a protein from the matrix; antibody (immunoglobulin produced by B lymphocytes in response to an antigen, binds it or related structures, e.g. epitopes); antigen; allergen — antigen inducing an immunoglobulin-E mediated allergic reaction (except gluten), of proteinaceous nature within the scope of this document; clone; monoclonal antibody (from a single hybridoma clone, to a single epitope) and polyclonal antibodies (several B cells, different epitopes of the same antigen); specificity of an antibody; cross-reactivity — binding of an antibody to substances other than the antigen of primary interest; epitope (linear or conformational); conjugate (antibody or other substance linked to an enzyme, fluorochrome, particle, radioactive label; enzyme conjugates require a substrate); western blot (transfer of electrophoretically separated proteins to a membrane and visualisation with a conjugate — not further discussed in the document); ELISA — in vitro assay combining enzyme-linked antibodies (or antigen) and a substrate giving a coloured or fluorescent product, qualitative or quantitative, usually in microwell plates.

STATUS (as of 03.10.2026). The PKN search engine (query “PN-EN 15633-1”, 03.10.2026) lists PN-EN 15633-1:2020-03 in Polish and English versions, “Introduces: EN 15633-1:2019 [IDT]”, without a withdrawal note; PN-EN 15633-1:2009 (English version) — “Withdrawn and replaced by PN-EN 15633-1:2020-03”.

HOW MANY LABORATORIES AND IN WHAT FORM (copy of the accreditation scope database, data up to 17.09.2026, read 03.10.2026). The number 15633-1 appears in 3 records at 2 laboratories: AB 381 (1) and AB 819 (2). In the current PCA documents (read 03.10.2026; AB 381 issue no. 29 of 10.06.2026, AB 819 issue no. 23 of 04.08.2026) the number appears at the same 2 laboratories; in every document the number of page markers equals the number of PDF pages, without repetitions; a search of all PCA documents downloaded from BIP on 03.10.2026 found no laboratories outside the database copy. The form “PN-EN 15633-1:2020-03” at AB 819, “PN-EN 15633-1” without year at AB 381 — in both together with kit manufacturers’ instructions (at AB 381 also with procedure PB/PACH/39). None of these items is suspended. The “Laboratories” tab (query “PN-EN 15633-1”) shows both laboratories — checked by a tab query on the labcoda.pl production server on 03.10.2026.

WHAT THE LABORATORIES TEST (items in PCA). AB 381: foodstuffs and environmental samples from food production and distribution areas — content of allergenic proteins by ELISA, according to manufacturers’ instructions. AB 819: a wide range of foods (meat, fish, ready meals, sweets, milk, seafood, fruit and vegetables, processed products, cereals, eggs and others) — gliadin (2.5–40) mg/kg with gluten by calculation, according to the RIDASCREEN Gliadin instructions of 03.06.2024; meat and meat products, ready meals — soy protein (2.5–6000) mg/kg, according to the RIDASCREEN FAST Soya instructions of 01.10.2025; ELISA.

WHERE A SEEMINGLY CORRECT RESULT IS EASY TO GET. Gluten: the definition of allergen in this standard excludes gluten (3.2.3) — determining gliadin with reference to PN-EN 15633-1 (AB 819) relies on the framework for immunological methods, but gluten is not an “allergen” in the sense of this definition. Processing: denaturation changes the antigenic properties of a protein, and cross-linkage its extractability from the matrix (3.1.1, 3.1.2) — a processed sample may give a low result although the ingredient is present. Cross-reactivity: an antibody can bind similar epitopes of other antigens (3.2.8) — a positive result does not prove the presence of exactly this ingredient. Type of antibodies: monoclonal ones recognize one epitope, polyclonal ones different epitopes (3.2.5, 3.2.6), which affects whether a processed protein is detected. Procedure: the standard is a framework — the specific extraction, calibration and range are given by the kit manufacturer’s instructions, listed next to the standard in PCA. Edition: PN-EN 15633-1:2009 is withdrawn and replaced; an item without year (AB 381) does not indicate the edition.

WHAT WE DO NOT GIVE. We do not quote the requirements for laboratories, procedure, validation criteria, interpretation of results or influencing factors of Clauses 4–7 — they lie outside the sample. We do not summarize EN 15842 or the kit manufacturers’ instructions in this entry. We do not give allergen labelling thresholds or gluten limit values.

Method principle

Protein extracted from the food sample is tested with an assay based on antibodies directed against a protein or peptide that is a marker of the allergenic ingredient. In an ELISA, an enzyme-linked antibody (or antigen) binds the analyte, and the added substrate gives a coloured or fluorescent product whose signal indicates the presence or amount of the marker. The standard sets the framework and criteria for such methods; the specific extraction and calibration are given by the procedure of the particular test.

Applications

• Qualitative detection and quantitative determination of food allergens and allergenic ingredients in foodstuffs (1)
• Antibody-based methods: ELISA and other immunoassays, including on-site testing (1, 3.3.2)
• Foodstuffs and environmental samples from food production — allergenic proteins, gliadin, soy protein — 3 accreditation scope records of 2 laboratories in PCA

Key parameters

Standard

Standard number

PN-EN 15633-1:2020-03

Title (PL)

Artykuły żywnościowe — Wykrywanie alergenów pokarmowych metodami immunologicznymi — Część 1: Postanowienia ogólne

Title (EN)

Foodstuffs — Detection of food allergens by immunological methods — Part 1: General considerations

Step-by-step procedure

1. Choice of method

   Antibody-based test against the marker protein; EN 15842 and PN-EN 15633-1 (1, 2). PN-EN 15633-1:2020-03 — 2009 edition withdrawn and replaced (PKN, 03.10.2026).

2. Protein extraction

   The extraction procedure is part of each method (introduction).

3. Determination

   Qualitative or quantitative ELISA (3.3.2; Clause 5 outside the sample).

4. Interpretation

   Expression of results and validation criteria (Clause 6 outside the sample).

5. Influencing factors

   Denaturation, cross-linkage, cross-reactivity (3.1, 3.2.8; Clause 7 outside the sample).

Required equipment and apparatus

Reagents, media and consumables

Health and safety (OHS)

• The part of the standard read contains no safety requirements; rules for handling reagents are given by the kit manufacturer’s instructions

Frequently asked questions