Anti-urease immunoglobulin (IgY) from egg yolk prevents Helicobacter pylori infection in a mouse model

Figures & data

Table 1. Urease activity after anti-urease IgY treatment.

Protein	Activity (mU/mg protein)
Urease A	7.54 mU/mg protein
Urease B	6.46 mU/mg protein

Figure 1. Confirmation of recombinant protein expression (UreA). Notes: Results of SDS-PAGE and western-blot analysis of Urease A. pET32a-UreaseA: 51 kDa. SDS-PAGE, Lane 1,3- Before induction; 2,4- Before induction.

Figure 2. Confirmation of recombinant protein expression (UreB). Notes: Results of SDS-PAGE and Western-blot analysis of Urease B protein. pET32a-UreaseB 81 kDa. SDS-PAGE, Lane 1,3- Before IPGT induction; 2,4- After IIPGT induction.

Figure 3. Detection of egg yolk antibody. Notes: Results of egg yolk antibody detection test by 12% SDS-PAGE.

Figure 4. Binding identification test for anti-bacteria. Notes: Binding affinity test for egg yolk antibody by 12% SDS-PAGE. M: Protein Marker, 1: H. pylori protein, 2: Anti-H against H. pylori. H. pylori-IgY Binding affinity was confirmed by western-blot.

Figure 5. Coagulation reaction confirmation test result. Notes: Cohesion test results. IgY sample 40 mg/ml starting, 2-fold dilution, optical microscopy at 200× magnification.

Table 2. Test results for coagulation by IgY concentration.

Concentration (mg/ml)	0	0.009	0.019	0.039	0.078	0.156	0.313	0.625	1.25	2.5	5	10	20	40
Aggregation	x	x	x	x	x	x	o	o	o	o	o	o	o	o

Figure 6. Urease inhibition result. Notes: Urease activity inhibition confirmation test results. *p < 0.05, **p < 0.01, ***p < 0.001 vs control.

Table 3. Test animal weight change results.

Group		Weeks
		0	1	2	3	4
G1	Normal	18.75 ± 0.70	19.68 ± 0.98	20.91 ± 0.75	22.28 ± 0.77	20.12 ± 0.89
G2	H. pylori only	18.78 ± 0.62	19.48 ± 0.82	20.09 ± 0.70	21.71 ± 1.03	22.83 ± 0.86
G3	H. pylori + Normal egg yolk powder 500 mg/kg	18.80 ± 0.65	19.45 ± 0.60	20.57 ± 0.71	22.20 ± 1.55	22.56 ± 0.97
G4	H. pylori + Anti-Urease IgY 50 mg/kg	18.72 ± 0.46	19.16 ± 0.60	20.19 ± 0.60	20.41 ± 0.66	20.88 ± 0.80
G5	H. pylori + Anti-Urease IgY 100 mg/kg	18.78 ± 0.69	19.96 ± 0.47	20.37 ± 0.77	21.90 ± 1.08	22.59 ± 0.98
G6	H. pylori + Anti-Urease IgY 250 mg/kg	18.58 ± 0.63	19.08 ± 0.97	20.56 ± 0.76	22.09 ± 0.97	22.83 ± 1.21
G7	H. pylori + Anti-Urease IgY 500 mg/kg	18.73 ± 0.37	19.05 ± 0.92	20.94 ± 0.76	22.11 ± 1.11	22.34 ± 1.13

Table 4. Reactivity in CLO Kit of H pylori infected mice gastric mucosa after treatment with anti-urease IgY.

Group	1	2	3	4	5	6	7	8	9	10	Positive ratio
Normal	●	●	◐	○	○	○	○	○	○	○	2/10
H. pylori only	●	●	●	●	●	●	●	●	●	●	10/10
H. pylori + Normal egg yolk powder 500 mg/kg	●	●	●	●	●	●	●	●	○	○	8/10
H. pylori + Anti-Urease IgY 50 mg/kg	●	●	●	●	●	○	○	○	○	○	5/10
H. pylori + Anti-Urease IgY 100 mg/kg	●	●	●	◐	○	○	○	○	○	○	3/10
H. pylori + Anti-Urease IgY 250 mg/kg	●	●	●	◐	○	○	○	○	○	○	3/10
H. pylori + Anti-Urease IgY 500 mg/kg	●	●	◐	◐	○	○	○	○	○	○	2/10

Notes: ○: negative, ◐: partially positive, ●: positive.

Table 5. Identification of H pylori through culture of gastric mucosa from infected mice after treatment with anti-urease IgY.

Group	1	2	3	4	5	6	7	8	9	10	Positive ratio
Normal	+	+	−	−	−	−	−	−	−	−	2/10
H. pylori only	+	+	+	+	+	+	+	+	+	+	10/10
H. pylori + Normal egg yolk powder 500 mg/kg	+	+	+	+	+	+	+	+	−	−	9/10
H. pylori + Anti-Urease IgY, 50 mg/kg	+	+	+	+	+	−	−	−	−	−	5/10
H. pylori + Anti-Urease IgY, 100 mg/kg	+	+	+	−	−	−	−	−	−	−	3/10
H. pylori + Anti-Urease IgY, 250 mg/kg	+	+	+	−	−	−	−	−	−	−	3/10
H. pylori + Anti-Urease IgY, 500 mg/kg	+	+	−	−	−	−	−	−	−	−	2/10