Isolation of Allergenically Active Cytochrome c from Kentucky Blue Grass Pollen

Abstract

An allergen, designated as Kentucky blue grass KBG-cytochrome c, was isolated from the nondialysable constituents (R) of the aqueous extract of KBG pollen by a combination of gel filtration and preparative isoelectrofocussing techniques. The allergen had an absorption spectrum characteristic of heme protein and had a molecular weight corresponding to 12,000 daltons. It was a basic protein with a pI value of 9.9 and contained all the common amino acids. KBG-cytochrome c elicited immediate hypersensitivity cutaneous reactions in individuals allergic to KBG pollen.

Monoclonal Antibodies to Proteins from Cocksfoot Grass <i>(Dactylis glomerata)</i> Pollen: Isolation and N-Terminal Sequence of a Major Allergen

“…The purification of allergic components from pollen extracts has been achieved by standard physicochemical separation techniques [1][2][3][4][5] and the use of immobilised monoclonal antibod ies (Mabs) as immunosorbents [6][7][8]. The availability of allergens in pure form has facilitated the determi nation of several partial N-terminal sequences [9][10][11] and a number of complete protein sequences [12][13][14] by Edman degradation.…”

Isolation and Partial Characterization of Allergen C from Kentucky Blue Grass Pollen

“…Thus, it may be said that Allergen C con sisted of one antigenic component. In a par allel study [7] an allergenically active cyto chrome c isolated from KBG pollen pos sessed certain properties such as color, size and a pi value that were similar to those of Allergen C. These comparisons made in ret rospect, suggest that Allergen C may have been contaminated with cytochrome c. However, the absorption spectrum charac teristic for cytochrome c was not detected in the preparation of Allergen C. The contami nation, if any, of Allergen C by cytochrome c would have to be at a level below that de tectable by the absorption study. The immu nological relationships between the two al lergens must await the availability of appro priate antisera to KBG cytochrome c.…”

Isolation of a Kentucky Blue Grass Pollen Allergen Using a Murine Monoclonal Antibody Immunosorbent

“…One hundred milligrams of KBG-R was fractionated by prepar ative isoelectrofocusing as described previously [9], Briefly, Ultrodex (LKB-Products AB, Stockholm-Bromma, Sweden) was swollen in a solution of KBG-R containing ampholyte buffer, pH range 3.5-10 (LKB). The gel was poured onto a horizontal glass tray (I I x 30 x 0.8 cm, Bio-Rad).…”