In-Vitro Simulated Gastric Fluid Digestion and Immunogenicity of Different Crustacean Protein Extracts

Figures & data

FIGURE 1 SDS-PAGE/Coomassie blue-staining of seven crustaceans protein extracts with same protein concentration. M, molecular weight proteins; (1) White-leg shrimp (Penaeus vannamei); (2) Female Chinese shrimp (Penaeus chinensis); (3) Male Chinese shrimp (Penaeus chinensis); (4) Southern rough shrimp (Trachypenaeus curvirostris); (5) Acetes chinensis; (6) Mantis shrimp (Squilla oratoria); (7) Crawfish (Procambarus clarkii).

TABLE 1 The molecular mass (kDa) of western blotting bands of seven crustacean protein extracts to 10 crustacean allergy patients sera

Crustacean varieties
	White-leg shrimp	Female Chinese shrimp	Male Chinese shrimp	Southern rough shrimp	Acetes chinensis	Mantis shrimp	Crawfish
Patients sera
A	38, 61, 42	61	37, 42, 81*	37	38	38	38*
B	38*	36, 42, 61, 83	37*, 42, 69*, 81*	37*	34, 38	38	38*
C	23, 38, 42, 61	23, 42, 61, 83*	23, 37, 42, 69*, 81*	25, 37	38	38	38
D	23, 38, 61	23, 42	23, 37*, 42*, 69*, 81*	25, 37	−	−	−
E	23, 38*	23, 42, 61	23, 37*, 42, 52, 69, 81*	25, 37	−	−	−
F	38	−	23, 37*, 42*, 69*, 81*	37	−	−	−
G	23, 38	23, 42, 61, 83	23, 37*, 42*, 69*, 81*	25, 37	34	26,54	38
H	23, 38, 42, 61	23, 42	23, 37*	25, 37*	38, 42	38	38*
I	23, 38*, 42, 61	23, 42, 83	37*, 42, 69, 81*	25, 37*	38	26, 38	38*
J	38*, 42	23, 34, 42, 61	23, 37*, 42, 69	25, 37*	34, 38	26, 30, 38*	38*

*stands for the high binding capacity with patients’ sera IgE;

A,B,C, … J, 10 crustacean allergic patients sera.

FIGURE 2 Western blotting detection of the crustacean protein extracts from seven different varieties. Samples were separated by SDS-PAGE followed by immunological detection using serum of 10 crustacean allergic patients (A,B,C, … J). M, molecular weight proteins; (1) White-leg shrimp (Penaeus vannamei); (2) Female Chinese shrimp (Penaeus chinensis); (3) Male Chinese shrimp (Penaeus chinensis); (4) Southern rough shrimp (Trachypenaeus curvirostris); (5) Acetes chinensis; (6) Mantis shrimp (Squilla oratoria); (7) Crawfish (Procambarus clarkii).

FIGURE 3 Indirect ELISA for tropomyosin in crustacean protein extracts.

FIGURE 4 SDS-PAGE analysis of SGF digestion on crustacean protein extracts. In the control experiments (Con) pepsin was excluded. The digest was withdrawn at different time intervals (0.5, 1, 2, 5, 10, 15, 30, and 60 min). M, molecular weight proteins; Con, the eatracts of no pepsin; (A) White-leg shrimp (Penaeus vannamei); (B) female Chinese shrimp (Penaeus chinensis); (C) male Chinese shrimp(Penaeus chinensis); (D) Southern rough shrimp (Trachypenaeus curvirostris); (E) Acetes chinensis; (F) Mantis shrimp (Squilla oratoria); (G) Crawfish (Procambarus clarkii).

FIGURE 5 IgE-Western blot (B) using rabbit polyclonal antibodies to detect the IgE-binding of crustacean protein extracts after digestion by SGF. The digest was withdrawn at different time intervals (0.5, 1, 2, 5, 10, 15, 30, and 60 min). M, molecular weight proteins; (1) White shrimp (Penaeus vannamei); (2) female Chinese shrimp (Penaeus chinensis); (3) male Chinese shrimp (Penaeus chinensis); (4) Southern rough shrimp (Trachypenaeus curvirostris); (5) Acetes chinensis; (6) Mantis shrimp (Squilla oratoria); (7) Crawfish (Procambarus clarkii).