• 한국축산식품학회지
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• 제25권2호
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• pp.250-256
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• 2005

To investigate the effect of bovine colostral whey fractions on in vitro proliferation of mouse splenocytes, polypeptide fractions were separated from acid whey into 3 fractions depending on molecular weight by ultrafiltration: Fraction I, which contains the polypeptide larger than 10,000 Da, Fraction n, which contains the polypeptide ranging from 1,000 Da to 10,000 Da and Fraction III, which contains the polypeptide smaller than 1,000 Da. Fraction II showed the highest proliferative effect of mouse splenocytes among the colostral whey fractions and this proliferative activity increased in dose dependent manner. Unheated Fraction II and Fraction III showed significantly (p<0.01) higher proliferative effects than others but heated Fraction II showed the highest enhancing effect of mouse splenocyte among heated whey fractions (p<0.01). The supplementation of Fraction II and Fraction m showed greater proliferative effect of mouse splenocytes stimulated by concanavalin A (Con A) than that of whole whey or Fraction L Proliferative effect of mouse splenocytes stimulated by phytohemagglutinin (PHA) was the highest when Fraction II was supplemented Proliferative effect of the colostral whey fractions on mouse splenocytes by stimulation of lipopolysaccharide (LPS) was markedly enhanced by supplementation of Fraction II and Fraction m compared with whole whey and Fraction L It was estimated that colostral whey fraction containing IGF-I positively affected proliferation of mouse splenocyte.

• Asian-Australasian Journal of Animal Sciences
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• 제19권2호
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• pp.297-304
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• 2006

Insulin-like growth factor-I (IGF-I) rich fraction, collected components between 1 kDa and 30 kDa, was fractionated from bovine colostral whey using an ultrafiltration membrane. IGF-I was confirmed in the collected IGF-I rich fraction by both SDS-PAGE and Western blotting. The concentration of IGF-I in the IGF-I rich fraction was 10 ng/mg protein. One hundred microliters of the reconstituted IGF-I rich fraction was intraperitoneally injected into ICR male mice for 2 weeks at 24 h intervals. The functions of peritoneal macrophages, including phagocytosis, interleukin (IL)-6 and tumor necrosis factor (TNF)-${\alpha}$ production, and nitric oxide and hydrogen peroxide production, were enhanced significantly by the administration of the IGF-I rich fraction in a dose-dependent manner (p<0.01). The proliferation of Concanavalin (Con) A-stimulated and Lipopolysaccharide (LPS)-stimulated splenocytes was also determined to have been enhanced significantly by the administration of the IGF-I rich fraction in a dose-dependent manner (p<0.01). Our results indicate that the administration of IGF-I rich fraction obtained from bovine colostral whey enhances both innate and acquired immunity for ICR male mice.

• 한국축산식품학회지
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• 제30권6호
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• pp.923-929
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• 2010

Colostral whey prepared from colostrum (pooled from first six post-partum milkings) was heated for 10 min at $100^{\circ}C$ Heated colostral whey was incubated with 1% enzymes (protein equivalent basis) for 15, 30, 60, 90, and 120 min at $50^{\circ}C$. Papain, pepsin, trypsin, and alcalase produced different degrees of hydrolysis (DH), 10.66%, 12.42%, 10.83%, and 25.31%, respectively, at an incubation time of 120 min. The SDS-PAGE reveals that significant amounts of bovine serum albumin (BSA), ${\beta}$-lactoglobulin (${\beta}$-LG), and ${\alpha}$-lactalbumin (${\alpha}$-LA) survived papain digestion. In contrast, pepsin completely removed BSA but not ${\beta}$-LG present in heated colostral whey. Alcalase completely eliminated BSA, ${\beta}$-LG, and ${\alpha}$-LA. This differential hydrolysis was confirmed by reversed-phase HPLC analysis. Using ion-exchange chromatography, fraction-1 (F-1) was obtained from alcalase hydrolysate at a NaCl gradient concentration of 0.25 M. Reversed-phase HPLC chromatograms of alcalase F-1 showed numerous small peaks, which probably indicate that a variety of new peptides were produced. Iron content of alcalase F-1 was 28.94 ppm, which was the highest among all enzyme fractions, whereas iron content of colostral whey was 36.56 ppm. Main amino acids contained in alcalase F-1 were Thr (15.45%), Glu (14.12%), and Ser (10.39%). Therefore, alcalase can be used to generate good iron-binding peptides in heated colostral whey, and the resulting iron-binding peptides could be suitable as a value-added food ingredient for food supplements.

• 한국축산식품학회지
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• 제26권1호
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• pp.113-120
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• 2006

젖소 초유 whey 내에 존재하는 IGF-I을 분리하기 위하여 ultrafiltration을 사용하여 30과 1 kDa 사이의 IGF-I rich fraction을 분획하였다. 분획한 IGF-I rich fraction내에 IGF-I의 존재 여부는 SDS-PACE와 western blotting으로 확인하였고, sandwich ELISA로써 분획 내 IGF-I을 정량하였다. 그 결과 IGF-I은 단백질 mg당 10 ng으로 측정되었다. 분획한 IGF-I rich fraction을 마우스 복강 macrophge의 면역 활성에 미치는 영향 즉, IL-6, NO, $TNF-{\alpha}$, Phagocytosis, $H_{2}O_{2}$,의 분비 유도 및 생성량을 측정하였고, splenocyte의 면역 활성으로는 splenocyte의 증식 효과와 natural killer cell의 항암작용을 측정하였다. 실험 결과 IGF-I rich fraction $1{\mu}g/mL$ 투여군에서 IL-6의 생성량은 9.85 ng, NO의 생성량은 $17.1{\mu}M$, phagocytosis는 양성 대조구에 대비하여 78.3%의 탐식작용을 나타내었고 $TNF-{\alpha}$와 $H_{2}O_{2}$의 생성량은 대조구에 대비하여 각각 34.5 및 6% 더 많은 양을 생산하였다. Splenocyte 면역활성에 미치는 영향으로 T cell과 B cell 증식은 대조군에 대비하여 각각 36 및 76%로 더 높은 증식 효과를 나타내었고, NK cell의 활성은 대조군보다 55.4%의 높은 활성을 보였다.

• 한국축산식품학회지
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• 제25권2호
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• pp.244-249
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• 2005

To investigate the effect of Holstein colostrum peptide fraction on proliferation of immune cell, polypeptide fractions were separated from acid whey into 3 fractions depending on molecular weight by ultrafiltration: Fraction I, which contains the polypeptide larger than 10,000 Da, Fraction n, which contains the polypeptide ranging from 1,000 Da to 10,000 Da and Fraction III, which contains the polypeptide smaller than 1,000 Da. EL-4 cell (murine T lymphoma cell) was used to evaluate immune enhancing effect of each fraction from Holstein colostrum. Fraction n showed the highest proliferative effect of the colostrum whey fractions on EL-4 cell at 1mg/mL compared with whole whey and other fractions and this proliferative activity was shown in dose dependent manner. Fraction n showed the highest proliferative effect on PMA (Phorbol 12-myristate 13-acetate) stimulated EL-4 cell. Heated Fraction n showed similar effect to native one on proliferation of both EL-4 cell and PMA stimulated EL-4 cell.

• 한국식품과학회지
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• 제34권4호
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• pp.694-699
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• 2002

본 연구에서는 분만 후 5일 이내에 분비되는 젖소 초유의 whey 및 whey 분획이 Th1 cell의 증식에 어떤 영향을 미치는지, 그리고 Th1 cell 증식에 직접적으로 관여하는 whey 분획이 macrophage의 $TNF-{\alpha}$ 분비에 미치는 영향을 조사하였다. 초유 whey를 ultrafiltration으로 분자량별로 분획한 결과, 단백질 성분의 회수율은 Fr. I, II, III가 각각 72%, 17.7%, 10.2%였으며, 당 성분의 회수율은 Fr. I, II, III가 각각 22.1%, 7.4%, 70.1%였다. Fr. II를 재분획한 Fr. P의 단백질 회수율과 Fr. O의 당 회수율은 각각 86.9%, 88.8%였다. 각각의 whey 분획의 농도 대비 Th1 cell 증식 효과를 검증한 결과, 1 mg/mL 농도에서 Fr. II가 Th1 cell을 가장 많이 자극시켰으며, 세포증식율은 67.1%였으나, Fr. II의 단백질 및 올리고당 분획의 세포증식효과는 없는 것으로 나타났다. Whey의 각각의 분획을 이용하여 $TNF-{\alpha}$ 분비 능력을 조사한 결과, Fr. O가 양성대조구로 사용한 LPS보다 약 80% 이상의 $TNF-{\alpha}$분비 유도 능력이 있는 것으로 나타났다.

• 한국축산식품학회지
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• 제24권2호
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• pp.171-175
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• 2004

젖소 초유 유청을 ultrafiltration으로서 30kDa 사이의 IGF-I rich fraction을 분획하였다. 분획한 IGF-I rich fraction은 SDS-PAGE와 Western blotting으로 IGF-I이 존재하는 것을 확인하였고, sandwich ELISA로써 그 함량을 측정한 결과 fraction중 IGF-I의 함량은 단백질 mg당 10ng으로 나타났다. IGF-I rich fraction으로 IEC-6, Detroit 551, EL-4 및 L6 in vitro세포의 증식에 미치는 효과를 실험한 결과 IEC-6 cell은 IGF-I 기준으로 10ng, 1 ng, 0.1ng, 0.01ng에서 각각 대조구에 비해서 60%, 53%, 30 그리고 20%의 세포증식효과를 나타내었다. IEC-6의 증식은 ICF-I의 투여량이 증가할수록 세포의 증식효과가 더 높게 나타났다. Detroit 551 cell은 10ng과 1ng수준에서 각각 56%와 26%의 세포증식 효과를 나타내었다. 그리고 EL-4 cell과 L6 cell은 10ng 수준에서 각각 53%와 46%의 세포증식 효과를 나타내었다. 모든 세포주에서 IGF-I 함유농도가 10 ng 투여구에서 세포 증식율이 가장 높았으며 IEC-6 cell, Detroit 551 및 EL-4 cell은 모두 약 60%의 세포 성장률을 나타내었고 L6 cell은 약 50%의 성장률을 보였다.