Anti ccd absorbent

1:20 Anti-CCD absorbent (EUROIMMUN Medizinische Labordiagnostika AG, Lübeck, SH, Germany) was added to the samples to be tested, and then mixed well and incubated at room temperature (18–25°C) in a shaker for 1 h. Immediately afterward, sIgE of mugwort and allergen components (Art v 1, Art v 3 and Art v 4), ragweed and allergen components (Amb a 1), bermuda grass and allergen components (Cyn d 1 and Cyn d 12), and timothy grass and allergen components (Phl p 1, Phl p 4, Phl p 5, Phl p 7, and Phl p 12), and CCD were tested via the EUROBlotMaster system. The inhibition rates of CCD on pollen allergens were calculated by (sIgE after CCD inhibition minus the corresponding values before CCD inhibition and then divide by before levels) (20 (link), 21 (link)).

Serum sIgE‐levels to honeybee venom (HBV), wasp venom (WV), hornet venom (HV), and recombinant allergens: rApi m 1, rApi m 2, icarapin (rApi m 10) from HBV, rVes v 5, phospholipase A1 (rVes v 1) from WV, and CCD were measured by Immunoblot (Euroimmun, Germany, Euroline DPA‐Dx insect venoms 3). Serum samples of 29/71 multiple‐positive patients were tested in the EUROLINE assay using the anti‐CCD Absorbent (Euroimmun). The absorbent (CCD‐inhibitor) for eliminating CCD‐sIgE was composed of glycoprotein bromelain extracted from pineapple. Serum samples were incubated with anti‐CCD Absorbent for 60 min at room temperature according to the manufacturer's instructions. The sera were retested using the same insect venoms profile, as previously mentioned.
The cut‐off value for positive sIgE‐test results was 0.35 kUA/L. The sIgE‐reactivity was categorized quantitatively into six classes: Class 1 (≥0.35 to <0.70 kUA/L), Class 2 (≥0.70 to <3.50 kUA/L), Class 3 (≥3.50 to <17.50 kUA/L), Class 4 (≥17.50 to <50.00 kUA/L), Class 5 (≥50.00 to <100.00 kUA/L), and Class 6 (≥100.00 kUA/L).

The isolated extracts, protein fractions, and individual proteins were placed on a EUROLINE strip (EUROIMMUN, Lübeck, Germany) specially designed for this study by EUROIMMUN. Sensitization was assessed according to the standard manufacturer's instructions. The EUROLINE intensity units (EL) were evaluated using the EUROLineScan software and cross-reactive carbohydrate determinants (CCD) positive sera were reanalyzed after inhibition with Anti-CCD Absorbent (EUROIMMUN). An intensity of 3 (class 1) or higher was rated as positive.