Study on calcium alginate microcapsule embedding IgY and its release properties

doi:10.3969/j.issn.1004-1524.2022.12.17

Acta Agriculturae Zhejiangensis ›› 2022, Vol. 34 ›› Issue (12): 2740-2749.DOI: 10.3969/j.issn.1004-1524.2022.12.17

• Food Science • Previous Articles Next Articles

Study on calcium alginate microcapsule embedding IgY and its release properties

LU Wenjing¹(), CHEN Di¹, YE Qin¹, ZHANG Cen¹, RUAN Lingli¹^,², XIAO Chaogeng¹^,^*(), YAO Yunxin³

1. Institute of Food Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
2. College of Food Science and Technology, Yunnan Agricultural University, Kunming 650201, China
3. Zhejiang AGS Biotechnology Company Limited, Changxing 313100, Zhejiang, China

Received:2022-03-28 Online:2022-12-25 Published:2022-12-26
Contact: XIAO Chaogeng

Abstract

Abstract:

Egg yolk immunoglobulin (IgY) is a highly active oral antibody that is closely related to human health. In order to maximize the retention of IgY's biological activity and improve its stability, the natural polysaccharide sodium alginate was used as the main wall material, and the gel bead was prepared by the method of retination-coagulation bath embedding IgY. Taking the encapsulation efficiency and loading capacity as the main indicators, the optimal preparation process of IgY-calcium sodium alginate gel beads was explored by the response surface method, and the in vitro gastrointestinal digestion model was used to evaluate the release effect of gel beads. The results showed that when sodium alginate was 28.30 g·L^-1, calcium chloride was 10.80 g·L^-1 and the mass ratio of core to wall was 0.46:1, the newly prepared IgY-calcium alginate gel beads had good molding effect, the encapsulation efficiency was (53.30±1.55)%, and the loading capacity was (10.90±0.26)%. Through macroscopic morphological observation, the gel bead particles had good roundness and uniformity, with a particle size of about (2.10±0.05) mm and a hardness of (4 396.87±331.62) g. After drying, the swelling ratio of the gel beads was 0.94±0.39 after 2 h in the simulated gastric juice, the cumulative release ratio of IgY was about (11.10±3.20)%, and after 2 h in the simulated intestinal fluid, the swelling ratio continued to increase to 4.81±0.32, and the cumulative release ratio of IgY reached (97.10±2.40)%. Therefore, IgY-calcium alginate gel beads had the effect of slowly releasing the core material in the gastric juice, and IgY could be basically released in the intestinal fluid.

Key words: egg yolk immunoglobulins, calcium alginate, sodium alginate, external release rate

CLC Number:

TQ460.4

LU Wenjing, CHEN Di, YE Qin, ZHANG Cen, RUAN Lingli, XIAO Chaogeng, YAO Yunxin. Study on calcium alginate microcapsule embedding IgY and its release properties[J]. Acta Agriculturae Zhejiangensis, 2022, 34(12): 2740-2749.

Figures/Tables 10

Table 1 Horizontal table of gel bead factors

水平 Level	海藻酸钠 Sodium alginate/ (g·L^-1)	氯化钙 Calcium chloride/ (g·L^-1)	芯壁比 Core-to-wall ratio
1	30	20	4∶6
0	25	15	3∶6
-1	20	10	2∶6

Fig.1 Influence of sodium alginate concentration on gel bead embedding effect Different lowercase letters in the figure indicated significant difference (P<0.05).

Fig.2 Influence of CaCl2 concentration on gel bead embedding effect

Fig.3 Influence of core-wall ratio on gel bead embedding effect

Table 2 Response surface test design and results of gel bead

试验号 Number	壁材浓度(X₁) Wall material concentration (X₁)/(g·L^-1)	氯化钙浓度(X₂) Calcium chloride concentration (X₂)/(g·L^-1)	芯壁比(X₃) Core-to-wall ratio (X₃)	包埋率 Encapsulation efficiency/%	载药量 Loading capacity/%
1	20	10	3∶6	29.55	8.79
2	30	15	4∶6	32.85	11.73
3	25	15	3∶6	51.37	15.29
4	20	20	3∶6	19.34	5.76
5	25	15	3∶6	50.91	15.15
6	30	15	2∶6	42.68	9.53
7	25	15	3∶6	53.87	16.03
8	25	15	3∶6	53.14	15.82
9	20	15	2∶6	7.55	1.67
10	30	20	3∶6	40.19	11.96
11	20	15	4∶6	33.53	11.79
12	25	10	4∶6	43.27	15.42
13	25	20	4∶6	49.85	17.76
14	25	10	2∶6	39.72	8.84
15	30	10	3∶6	54.27	16.15
16	25	20	2∶6	23.18	5.16
17	25	15	3∶6	37.63	11.20

Table 3 Variance analysis of regression model for gel bead embedding rate

方差来源 Source of variance	平方和 Sum of squares	自由度 Degree of freedom	均方 Mean square	F值 F value	P值 P value
模型Model	2 641.37	9	293.49	8.40	0.005 2
X₁	800.48	1	800.48	22.91	0.002 0
X₂	146.73	1	146.73	4.20	0.079 6
X₃	268.75	1	268.75	7.69	0.027 6
X₁X₂	3.78	1	3.78	0.11	0.751 9
X₁X₃	320.73	1	320.73	9.18	0.019 1
X₂X₃	133.60	1	133.60	3.82	0.091 4
$X_{1}^{2}$	576.69	1	576.69	16.51	0.004 8
$X_{2}^{2}$	14.37	1	14.37	0.41	0.541 8
$X_{3}^{2}$	306.42	1	306.42	8.77	0.021 1
残差Residual	244.54	7	34.93	—	—
失拟项Missing item	65.85	3	21.95	0.49	0.707 1
误差项Error term	178.70	4	44.67	—	—
总和Sum	2 885.92	16	—	—	—

Table 3 Variance analysis of regression model for gel bead embedding rate

方差来源 Source of variance	平方和 Sum of squares	自由度 Degree of freedom	均方 Mean square	F值 F value	P值 P value
模型Model	2 641.37	9	293.49	8.40	0.005 2
X₁	800.48	1	800.48	22.91	0.002 0
X₂	146.73	1	146.73	4.20	0.079 6
X₃	268.75	1	268.75	7.69	0.027 6
X₁X₂	3.78	1	3.78	0.11	0.751 9
X₁X₃	320.73	1	320.73	9.18	0.019 1
X₂X₃	133.60	1	133.60	3.82	0.091 4
$X_{1}^{2}$	576.69	1	576.69	16.51	0.004 8
$X_{2}^{2}$	14.37	1	14.37	0.41	0.541 8
$X_{3}^{2}$	306.42	1	306.42	8.77	0.021 1
残差Residual	244.54	7	34.93	—	—
失拟项Missing item	65.85	3	21.95	0.49	0.707 1
误差项Error term	178.70	4	44.67	—	—
总和Sum	2 885.92	16	—	—	—

Table 4 Variance analysis of regression model of gel bead load

方差来源 Source of variance	平方和 Sum of squares	自由度 Degree of freedom	均方 Mean square	F值 F value	P值 P value
模型Model	301.00	9	33.44	7.76	0.006 6
X₁	57.03	1	57.03	13.23	0.008 3
X₂	9.16	1	9.16	2.12	0.188 3
X₃	124.03	1	124.03	28.77	0.001 0
X₁X₂	0.34	1	0.34	0.078	0.788 0
X₁X₃	15.68	1	15.68	3.64	0.098 1
X₂X₃	9.06	1	9.06	2.10	0.190 4
$X_{1}^{2}$	53.78	1	53.78	12.48	0.009 6
$X_{2}^{2}$	0.89	1	0.89	0.21	0.663 8
$X_{3}^{2}$	25.15	1	25.15	5.83	0.046 4
残差Residual	30.18	7	4.31	—	—
失拟项Missing item	14.35	3	4.78	1.21	0.414 0
误差项Error term	15.82	4	3.96	—	—
总和Sum	331.18	16	—	—	—

Table 4 Variance analysis of regression model of gel bead load

方差来源 Source of variance	平方和 Sum of squares	自由度 Degree of freedom	均方 Mean square	F值 F value	P值 P value
模型Model	301.00	9	33.44	7.76	0.006 6
X₁	57.03	1	57.03	13.23	0.008 3
X₂	9.16	1	9.16	2.12	0.188 3
X₃	124.03	1	124.03	28.77	0.001 0
X₁X₂	0.34	1	0.34	0.078	0.788 0
X₁X₃	15.68	1	15.68	3.64	0.098 1
X₂X₃	9.06	1	9.06	2.10	0.190 4
$X_{1}^{2}$	53.78	1	53.78	12.48	0.009 6
$X_{2}^{2}$	0.89	1	0.89	0.21	0.663 8
$X_{3}^{2}$	25.15	1	25.15	5.83	0.046 4
残差Residual	30.18	7	4.31	—	—
失拟项Missing item	14.35	3	4.78	1.21	0.414 0
误差项Error term	15.82	4	3.96	—	—
总和Sum	331.18	16	—	—	—

Fig.4 Response surface diagram of the interaction of the various factors on the encapsulation efficiency and loading capacity of gel bead

Fig.5 Macro morphology of IgY-calcium alginate gel beads (×50 times)

Table 5 Swelling ratio and release ratio of IgY-calcium alginate gel beads during the gastrointestinal digestion phase

消化时间 Digestion time/h		溶胀度 Swelling ratio	释放率 Release ratio/%
胃Stomach	1	0.90±0.24	9.80±2.40
	2	0.94±0.39	11.10±3.20
肠Intestine	1	2.26±0.15	91.40±4.20
	2	4.81±0.32	97.10±2.40

References 29

[1]	POLSON A, VON WECHMAR M B, VAN REGENMORTEL M H. Isolation of viral IgY antibodies from yolks of immunized hens[J]. Knee Surgery, Sports Traumatology, Arthroscopy, 1980, 9(5): 475-493.
[2]	HAU J, HENDRIKSEN C F. Refinement of polyclonal antibody production by combining oral immunization of chickens with harvest of antibodies from the egg yolk[J]. ILAR Journal, 2005, 46(3): 294-299. PMID
[3]	LARSSON A, CARLANDER D, WILHELMSSON M. Antibody response in laying hens with small amounts of antigen[J]. Food and Agricultural Immunology, 1998, 10(1): 29-36. DOI URL
[4]	CARLANDER D, KOLLBERG H, WEJÅKER P E, et al. Peroral immunotherapy with yolk antibodies for the prevention and treatment of enteric infections[J]. Immunologic Research, 2000, 21(1): 1-6. PMID
[5]	张鹏飞, 林花荣, 陈功义. 口服免疫球蛋白对动物健康影响的研究进展[J]. 中国饲料, 2019(22): 20-23.
	ZHANG P F, LIN H R, CHEN G Y. Research progress on the effects of oral immunoglobulin in animal health[J]. China Feed, 2019(22): 20-23. (in Chinese with English abstract)
[6]	HEO C S, LEE J J, BAEK Y J, et al. Food containing active strains for inhibiting infection and treating gastritis, gastric and duodenal ulcers: US6491956 B2[P/OL]. 2002-12-10[2022-03-27]. https://www.freepatentsonline.com/6491956.html.
[7]	张璐, 刘珍珍, 宁长申, 等. IgY在免疫检测及治疗中的应用进展[J]. 动物医学进展, 2017, 38(1): 78-81.
	ZHANG L, LIU Z Z, NING C S, et al. Advance in IgY application on immunodetection and disease treatment[J]. Progress in Veterinary Medicine, 2017, 38(1): 78-81. (in Chinese with English abstract)
[8]	NAGARAJ S, RAMLAL S, KINGSTON J, et al. Development of IgY based sandwich ELISA for the detection of staphylococcal enterotoxin G (SEG), an egc toxin[J]. International Journal of Food Microbiology, 2016, 237: 136-141. DOI PMID
[9]	SUI J, CAO L, LIN H. Antibacterial activity of egg yolk antibody (IgY) against Listeria monocytogenes and preliminary evaluation of its potential for food preservation[J]. Journal of the Science of Food and Agriculture, 2011, 91(11): 1946-1950. DOI URL
[10]	HE J X, HU J J, THIRUMALAI D, et al. Development of indirect competitive ELISA using egg yolk-derived immunoglobulin (IgY) for the detection of Gentamicin residues[J]. Journal of Environmental Science and Health, Part B, 2016, 51(1): 8-13. DOI URL
[11]	曹侃, 李双双. 微胶囊技术在保健食品中的应用进展[J]. 食品安全导刊, 2019(28): 58-61.
	CAO K, LI S S. Application of microencapsulation technology in health food[J]. China Food Safety Magazine, 2019(28): 58-61. (in Chinese)
[12]	耿凤, 邵萌, 魏健, 等. 微胶囊技术在保护天然活性成分中的应用研究进展[J]. 食品与药品, 2020, 22(3): 250-255.
	GENG F, SHAO M, WEI J, et al. Progress in application of microcapsule technology in protection of natural active ingredients[J]. Food and Drug, 2020, 22(3): 250-255. (in Chinese with English abstract)
[13]	ARISANTI C I S, SUKAWATI C B A C, PRASETIA I G N J A, et al. Stability of anthocyanins encapsulated from purple sweet potato extract affected by maltodextrin concentration[J]. Macromolecular Symposia, 2020, 391(1): 1900127. DOI URL
[14]	刘丽英, 王圣洁. 海藻酸钙凝胶微球的制备和pH敏感释放[J]. 中国组织工程研究与临床康复, 2009, 13(42): 8303-8306.
	LIU L Y, WANG S J. Preparation of sodium alginate microspheres used for pH-sensitive drug delivery[J]. Journal of Clinical Rehabilitative Tissue Engineering Research, 2009, 13(42): 8303-8306. (in Chinese with English abstract)
[15]	陈凤莲, 刘文娟, 谢江, 等. 缓释定向释放型IgY微胶囊的制备及其效果评价[J]. 中国生物制品学杂志, 2016, 29(2): 181-187.
	CHEN F L, LIU W J, XIE J, et al. Preparation and effect evaluation of directional and sustainedreleased IgY microcapsules[J]. Chinese Journal of Biologicals, 2016, 29(2): 181-187. (in Chinese with English abstract)
[16]	国家质量监督检验检疫总局. 出口乳、蛋、豆类食品中蛋白质含量的测定考马斯亮蓝法: SN/T 3926—2014[S]. 北京: 中国标准出版社, 2014.
[17]	谭文芳, 郭伦爱, 段文韬, 等. 海藻酸钠-壳聚糖P-VP8^*IgY微胶囊的制备及其释放性能[J]. 沈阳药科大学学报, 2018, 35(10): 825-830.
	TAN W F, GUO L A, DUAN W T, et al. Preparation of P-VP8^IgY-loaded sodium alginatechitosan microcapsules and evaluation of its release performance[J]. Journal of Shenyang Pharmaceutical University*, 2018, 35(10): 825-830. (in Chinese with English abstract)
[18]	包永华, 陆芝娟, 刘莉, 等. 响应面设计法优化海藻酸钙凝胶珠制备条件[J]. 食品工业, 2012, 33(7): 28-30.
	BAO Y H, LU Z J, LIU L, et al. Optimization of preparation conditions in producing calcium alginate bead gels using box-behnken[J]. The Food Industry, 2012, 33(7): 28-30. (in Chinese with English abstract)
[19]	李秀凉, 田丽爽, 张玉娟, 等. 响应面法优化海藻酸钙包囊的制备条件[J]. 黑龙江大学工程学报, 2012, 3(3): 62-66.
	LI X L, TIAN L S, ZHANG Y J, et al. Optimization of conditions for preparation of calcium alginate capsule by response surface method[J]. Journal of Engineering of Heilongjiang University, 2012, 3(3): 62-66. (in Chinese with English abstract)
[20]	DAVARCI F, TURAN D, OZCELIK B, et al. The influence of solution viscosities and surface tension on calcium-alginate microbead formation using dripping technique[J]. Food Hydrocolloids, 2017, 62: 119-127. DOI URL
[21]	KOVACS-NOLAN J, MINE Y. Microencapsulation for the gastric passage and controlled intestinal release of immunoglobulin Y[J]. Journal of Immunological Methods, 2005, 296(1/2): 199-209. DOI URL
[22]	张新荣. 抗体微胶囊制备及其保护效果研究[D]. 太谷: 山西农业大学, 2013.
	ZHANG X R. Preparation of IgG microcapsule and analysis of its protective effect[D]. Taigu: Shanxi Agricultural University, 2013. (in Chinese with English abstract)
[23]	LI X Y, JIN L J, UZONNA J E, et al. Chitosan-alginate microcapsules for oral delivery of egg yolk immunoglobulin (IgY): in vivo evaluation in a pig model of enteric colibacillosis[J]. Veterinary Immunology and Immunopathology, 2009, 129(1/2): 132-136. DOI URL
[24]	顾其胜. 海藻酸盐基生物医用材料与临床医学[M]. 上海: 上海科学技术出版社, 2015.
[25]	MOGHADAM H, SAMIMI M, SAMIMI A, et al. Electro-spray of high viscous liquids for producing mono-sized spherical alginate beads[J]. Particuology, 2008, 6(4): 271-275. DOI URL
[26]	李嘉颖. 海藻酸钠复合水凝胶包埋IgY及其释放特性研究[D]. 无锡: 江南大学, 2019.
	LI J Y. Encapsulation of IgY by sodium alginate-based composite hydrogels and study on its release properties[D]. Wuxi: Jiangnan University, 2019. (in Chinese with English abstract)
[27]	白菲菲, 黄金, 王冠华, 等. 叶黄素-海藻酸钠微球的制备及质量评价[J]. 中国食品添加剂, 2018(4): 93-98.
	BAI F F, HUANG J, WANG G H, et al. Preparation and quality evaluation of lutein-alginate microspheres[J]. China Food Additives, 2018(4): 93-98. (in Chinese with English abstract)
[28]	李椿方, 袁騉, 梁浩, 等. 锐孔-凝固浴法制备硫苷粗提物微胶囊的研究[J]. 食品研究与开发, 2016, 37(6): 107-111.
	LI C F, YUAN K, LIANG H, et al. Study of microcapsules containing glucosinolate crude extracts prepared by orifice-coagulation bath method[J]. Food Research and Development, 2016, 37(6): 107-111. (in Chinese with English abstract)
[29]	温霜, 巨晓洁, 谢锐, 等. 肠靶向海藻酸钙基微胶囊的制备及控释性能研究[J]. 化工学报, 2020, 71(8): 3797-3806.
	WEN S, JU X J, XIE R, et al. Fabrication and controlled-release properties of intestinal-targeted Ca-alginate-based capsules[J]. CIESC Journal, 2020, 71(8): 3797-3806. (in Chinese with English abstract)

Study on calcium alginate microcapsule embedding IgY and its release properties

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