JP4886983B2 - 抗体を含有する機能性組成物 - Google Patents
抗体を含有する機能性組成物 Download PDFInfo
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- JP4886983B2 JP4886983B2 JP2004357105A JP2004357105A JP4886983B2 JP 4886983 B2 JP4886983 B2 JP 4886983B2 JP 2004357105 A JP2004357105 A JP 2004357105A JP 2004357105 A JP2004357105 A JP 2004357105A JP 4886983 B2 JP4886983 B2 JP 4886983B2
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- antibody
- comparative example
- functional composition
- bacteria
- immunostimulatory
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- FYKDNWHPKQOZOT-UHFFFAOYSA-M sodium;dihydrogen phosphate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].OP(O)([O-])=O.OC(=O)CC(O)(C(O)=O)CC(O)=O FYKDNWHPKQOZOT-UHFFFAOYSA-M 0.000 description 1
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Landscapes
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Description
Korhonen H. et al. Bovine milk antibodies for health, British J. Nutrition,84,suppl.1,S135-S146 (2000) Bolke E, et al. Shock 17, 9-12 (2002) ウイリアムE.ポール編/多田富雄監訳、基礎免疫学(東京大学出版会)1986年、上181頁
1)抗体と免疫賦活物質を有効成分として含有する機能性組成物、
2)抗体が乳由来の抗体である請求項1に記載の機能性組成物、
3)免疫賦活物質が乳酸菌、ビフィズス菌以外の毒素を産生しない細菌の菌体または菌体成分である請求項1または2に記載の機能性組成物、
4)免疫賦活物質がカビ・酵母の菌体または菌体成分である請求項1または2に記載の機能性組成物、
5)免疫賦活物質がキノコ類の菌体または菌体成分である請求項1または2に記載の機能性組成物、
6)免疫賦活物質が細菌、カビ・酵母およびキノコ類から選ばれる菌類の菌体の酵素分解物である請求項1または2に記載の機能性組成物、および
7)免疫賦活物質がSOD(スーパーオキサイドディスムターゼ)である請求項1または2に記載の機能性組成物。
乳清タンパクあるいは抗体を含有する機能性組成物を100mg取り、50mlの50mMリン酸緩衝液(pH6.8)に溶解し、2時間以上撹拌してから、その10ml以上を0.45μメンブランフィルターでろ過した。一方、プロテインGカラム(Amersham、HiTrap Protein G HP 1ml)を50mMリン酸緩衝液(pH6.8)で平衡化し、前記メンブランフィルターでろ過した試料10mlをカラムに流し、同緩衝液(pH6.8)で洗浄後、100mMグリシン塩酸緩衝液(pH2.7)で抗体を溶出させた。
前記測定法に従い、市販の乳清タンパク中の抗体含量を測定したところ、Proliant8000(James Farrell & Co.製)、IgG含有濃縮乳清タンパク((株)アオテアロア)はそれぞれ50.0mg/g、166mg/gであった。
下記表1に示す配合(数値は質量比)で4種類の機能性組成物を作製した。表1には機能性組成物の抗体含量も併せて示した。また、比較例1としてProliant8000、比較例2として納豆菌の凍結乾燥菌体、比較例3としてアガリクス乾燥粉末、比較例4としてビール酵母粉末、比較例5としてカゼイン分解物、比較例6としてプロラミン処理SOD(オキシカイン(登録商標)、イゾセル社(仏)製)を以下の試験に使用した。さらに2種の免疫賦活物質の組成物を調製し、比較例7として使用した。
実施例1 実施例2 実施例3 実施例4 実施例5 比較例7Proliant8000 99 80 - 90 - -
IgG含有濃縮乳清タンパク - - 60 - 6 -
納豆菌(凍結乾燥菌体) 1 - - - - -
アガリクス乾燥粉末 - 20 - - - 50
ビール酵母粉末 - - 40 - - 50
プロラミン処理SOD - - - 10 - -
カゼイン分解物 - - - - 94 -
抗体含量(mg/g) 49.5 40.0 99.6 45.0 10.0 -
6週齡のBALB/c系雌マウス1群6匹に、実施例1〜5、比較例1〜7のそれぞれを1質量%加えた混餌飼料を与えて、2週間飼育した。また、対照例として何も添加しない飼料を与えて、同様に飼育した。その後、卵白アルブミン10μg、水酸化アルミニウム1mgを含む0.1mlの生理食塩水で免疫し、免疫の14日後に眼底静脈叢より採血し血清を分離した。この血清中の抗卵白アルブミンIgG抗体(抗EAIgG)濃度を次のように求めた。
実施例6で採取した血清中の抗卵白アルブミンIgE抗体(抗EAIgE)濃度を次のように求めた。すなわち、平底96穴マイクロプレート(Nunc、439454)に、PBSに溶解した抗マウスIgE抗体50μlを加え、4℃に一晩置いた。洗浄液(0.9%塩化ナトリウム、0.05%Tween20)で洗浄後、ウサギ血清200μlを加え、37℃、1時間置いてブロッキングした。洗浄後、1%ウサギ血清、0.05%Tween20、3%塩化ナトリウムを含むPBS(溶液B)にて希釈した上記血清を加え、37℃、1時間置いた。洗浄後、ビオチンを結合させた卵白アルブミンの溶液Bによる希釈液50μlを加え、37℃、1時間置いた。洗浄後、ペルオキシダーゼ標識アビジン(シグマ社製)の溶液Bによる2500倍希釈液50μlを加え、37℃、1時間置いた後、洗浄し、基質溶液(オルトフェニレンジアミン40mg、30%過酸化水素20μl/クエン酸−リン酸ナトリウム緩衝液100ml)100μlを加え、室温に置き、492nmにおける吸光度を測定した。その吸光度の値(各群の平均値±標準偏差)を表2に併せて示した。吸光度と抗体濃度とは正の相関があるため、吸光度が大きければ抗体濃度が高いことを表す。
抗EAIgG(μg/ml) 抗EAIgE(492nm吸光度)
対照例 242±87 0.713±0.203
比較例1 513±148 0.494±0.141
比較例2 480±137 0.543±0.163
比較例3 424±126 0.537±0.167
比較例4 446±128 0.584±0.194
比較例5 491±142 0.514±0.158
比較例6 452±138 0.553±0.170
実施例1 903±245 0.208±0.091
実施例2 874±230 0.213±0.118
実施例3 988±274 0.198±0.087
実施例4 790±189 0.234±0.133
実施例5 951±252 0.202±0.090
比較例7 587±176 0.402±0.139
被験者として、男女15名(21〜30歳:9名、31〜40歳:3名、41〜60歳:3名)を3つの群(各群5名)に分け、それぞれ比較例1、比較例2、実施例1の粉末を一日10g、100mlの水に懸濁させ、毎朝食後に飲用してもらった。2週間継続飲用してもらい、継続飲用前と継続飲用後に次のようにしてIL−12産生誘導の測定を行った。
被験者として、男女15名(21〜30歳:9名、31〜40歳:3名、41〜60歳:3名)を3つの群(各群5名)に分け、それぞれ比較例1、比較例2、実施例1の粉末を一日10g、100mlの水に懸濁させ、毎朝食後に飲用してもらった。2週間継続飲用してもらい、継続飲用中の後半の一週間について排便に関するアンケート調査を行った。なお、継続飲用前の一週間についても同様のアンケート調査を行っている。調査項目は(1)排便回数、(2)排便量、(3)糞便色調、(4)糞便臭気とし、結果(各群の平均値±標準偏差)を表3に示した。
(3)糞便色調は黄色:0、薄い黄土色:1、黄土色:2、茶色:3、こげ茶色:4、
(4)糞便臭気は、ほとんど気にならない:0、わずかに臭い:1、普通:2、臭い:3、ひどく臭い:4、で表した数値の平均値である。
継続飲用前 継続飲用中
(1)排便回数/週 比較例1 5.6±0.8 6.0±0.7
比較例2 5.5±0.8 5.9±0.7
実施例1 5.7±0.8 6.8±0.6
(2)排便量/週 比較例1 29.0±10.8 30.0±10.7
比較例2 29.5±10.5 29.9±10.7
実施例1 29.7±10.8 31.8±10.6
(3)糞便色調 比較例1 3.7±0.8 3.0±0.7
比較例2 3.5±0.8 2.9±0.7
実施例1 3.6±0.8 1.7±0.6
(4)糞便臭気 比較例1 2.8±0.6 1.4±0.5
比較例2 2.6±0.6 2.1±0.5
実施例1 2.8±0.6 0.4±0.3
Claims (3)
- 消化管の受動免疫物質と能動免疫物質とを含有し、免疫調整および整腸に使用するための経口投与用機能性組成物であって、
該受動免疫物質が乳由来の抗体であり、
該能動免疫物質が、(1)納豆菌の菌体または菌体成分、(2)酵母の菌体または菌体成分、(3)スーパーオキサイドディスムターゼからなる群から選択された少なくとも一種である、
経口投与用機能性組成物。 - 前記受動免疫物質を10mg/g以上含む、請求項1に記載の経口投与用機能性組成物。
- 前記能動免疫物質を1質量%以上含む、請求項1又は2に記載の経口投与用機能性組成物。
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