JP4509114B2 - 幼児用調製乳 - Google Patents
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- JP4509114B2 JP4509114B2 JP2006526518A JP2006526518A JP4509114B2 JP 4509114 B2 JP4509114 B2 JP 4509114B2 JP 2006526518 A JP2006526518 A JP 2006526518A JP 2006526518 A JP2006526518 A JP 2006526518A JP 4509114 B2 JP4509114 B2 JP 4509114B2
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- A—HUMAN NECESSITIES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
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Description
本願明細書において使用される用語「オステオポンチン」または「OPN」は、乳に由来するオステオポンチンのために使用し、乳中でタンパク質分解された天然に存在するOPNに由来の断片またはペプチド、または、WO 01/49741に提案されている方法によって得られる、スプライシングされた、リン酸化された、もしくはグリコシル化されたバリアントを含む。
ヒトの母乳サンプルを、分娩後10-58日の健康な10人の母親から回収した。乳を搾乳機によって回収し、各搾乳から2mlをサンプリングし、全ての日の乳を得るためにプールした。高速遠心することによって、全乳からクリームを抽出し、脱脂乳と分離した。脱脂乳を0.2ml画分に分注し、直接分析するか、必要なときまで-20℃で凍結した。
全ての乳サンプルにおけるタンパク質濃度を、スタンダードとしてウシ血清アルブミンを用いたブラッドフォード法によって求めた。製造者によって供給される説明書にしたがって、分析を行った。
個々の乳および市販の幼児用調製乳におけるOPNの濃度を、サンドイッチELISAによって測定した。このアッセイのために、ウシおよびヒトOPNに対する、ウシおよびヒトの乳から精製したネイティブなOPNに対する抗血清を、それぞれ、Dako(Glostrup, Denmark)においてウサギで調製した。ウシOPNは、原則的に、記載されているように(Sorensen ES, Petersen TE, 1993, Purification and characterization of three proteins isolated from the proteose peptone fraction of bovine milk, J. Dairy Res. 60: 189-197)、精製した。ヒト乳OPNは、原則的に、記載されているように(Senger DR, Perruzzi CA, Papadopoulos A, Tenen DG, 1989, Purification of a milk closely similar to tumor-secreted phosphoproteins and osteopontin. Biochem. Biophys. Acta 996: 43-48)、精製した。更に高純度とするために、ウサギへの免疫前に、両方のタンパク質を逆相クロマトグラフィーに供した。抗血清のIgG画分を、コンカバリンAアフィニティによって精製し、直接ELISAに用い、または、高い特異性の抗体を得るためにOPNアフィニティカラム上で精製した。抗体の特異性を、乳サンプルおよび精製OPNのウェスタンブロッティング分析によってチェックし、確認した。開発したELISAは、検出限界が<5ng/ml乳であり、10-300ng/mlの範囲で直線性を有するというように、非常に高感度であり、したがって、乳サンプルを分析前に、2000-23000倍希釈した。全てのサンプルを、測定ごとに6-12倍希釈して三サンプルずつ測定した。ELIAの信頼性を、既知の量のOPNをスパイクした乳サンプルによって確認したところ、これらの測定で使用される範囲においてはスパイクしたOPNの量が完全に検出でき、本アッセイは定量的であることが示された。同様に、ヒト乳サンプルからのOPNの精製によって、ELISAによって測定した濃度と一致するOPN収率が示された。
プールしたウシ乳のOPNのレベルを、4ヶ月間で四回測定した(表1)。OPN濃度の平均は、18.28mg/リットルであった。文献に従うと、総タンパク質は35,000mg/リットルであった。ウシ乳中のOPN含量は、総タンパク質の0.05%(w/w)である。
25-35歳(平均29歳)の10人の母親からの乳を、分娩後10-58日後にサンプリングし(分娩後平均21.6日)、OPNおよび総タンパク質を分析した。データを表2にまとめる。母乳のOPNは、22.44mg/リットルから257.35mg/リットルの範囲であり、平均は138.5mg/リットルであった。乳中の総タンパク質の濃度は、8327mg/リットルから13268mg/リットルの範囲であり、平均は10821mg/リットルであった。OPNは、平均して、ヒト乳中の総タンパク質の1.3%(w/w)を占め、一人の母親においては、乳タンパク質の3%を越えてOPNが存在した。
125g/リットルに基づき、mg OPN/リットル レディトゥフィード調製乳としてデータを計算した。
ヒトの腸細胞は、原則的に、記載されているように(Agnholt J, Kaltoft K, 2001. Infliximab downregulates interferon production in activated gut T-lymfocytes form patients with Crohn’s diseas. Cytokine. 15: 212-222)、入手し、IL-2およびIL-4存在下で培養した。IL-2およびIL-4の存在下での培養によって、受容体;TCRαβ+、CD4+CD45RO+の発現などの保存された細胞特性を有するT細胞の増殖が促進された。
100-300mg/リットルレベルのOPNは、以下の実施例における、幼児用調製乳のための適した量であると考えられる。しかしながら、本発明の範囲は、この量に制限するものではない。なぜならば、食料、ライフスタイル、地理、民族などが原因で、ヒト母乳組成物には変動が存在するからである。
<スターター調製乳>
スターター調製乳は、最初の4-6ヶ月内に必要な栄養素をカバーする赤ん坊のための食物を意味する。
情報源:新生児および赤ん坊のための幼児用調製乳およびサプリメントに関するAnnouncement no.202:
タンパク質含量=窒素含量×6.38(ウシ乳タンパク質である場合)
タンパク質含量=窒素含量×6.25(豆乳タンパク質単離物および部分的に加水分解したタンパク質である場合)
エネルギー:2200kJ/100g(525kcal/100g)
再構成:リットル当り125g
<EU内におけるタンパク質含有量のための規制要件>
情報源:新生児および赤ん坊のための幼児用調製乳およびサプリメントに関するAnnouncement no.202:
タンパク質含量=窒素含量×6.38(ウシ乳タンパク質である場合)
タンパク質含量=窒素含量×6.25(豆乳タンパク質単離物)
エネルギー:2170kJ/100g(520kcal/100g)
再構成:リットル当り150g
現在、この領域の規制はないが、現在進行中である。LBW赤ん坊のための幼児用調製乳の製造者は、現在、小児科医および「欧州連合におけるマーケティングのための、低出生体重調製乳の組成物に対するガイドラインについてのIDACE提案」から勧告を受けている。これらのガイドラインは、EUにおける来るべき法律のベースとなると予想されている。
タンパク質含量=窒素含量×6.38(ウシ乳タンパク質および加水分解ウシ乳タンパク質である場合)
エネルギー:315kJ/100ml(75kcal/100ml)
再構成:リットル当り150g
Claims (6)
- ウシの乳オステオポンチンで補充した幼児用調製乳またはベビーフードであって、前記乳オステオポンチンが、総タンパク質含量の1%から3%である、幼児用調製乳またはベビーフードであって、前記乳オステオポンチンが、レディトゥフィード調製乳中で少なくとも50-250mg/リットルとなる量で含まれている、幼児用調製乳またはベビーフード。
- 前記幼児用調製乳が、スターター調製乳である、請求項1に記載の幼児用調製乳またはベビーフード。
- 前記幼児用調製乳が、フォローオン調製乳である、請求項1または2に記載の幼児用調製乳またはベビーフード。
- 幼児用調製乳またはベビーフードにおけるサプリメントとしての、ウシの乳オステオポンチンの使用であって、前記乳オステオポンチンが、総タンパク質含量の1%から3%である、使用であって、前記乳オステオポンチンが、レディトゥフィード調製乳中で少なくとも50-250mg/リットルとなる量である、使用。
- 前記幼児用調製乳が、スターター調製乳である、請求項4に記載の使用。
- 前記幼児用調製乳が、フォローオン調製乳である、請求項4または5に記載の使用。
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AU2014286123B2 (en) * | 2013-07-05 | 2019-02-28 | Arla Foods Amba | Mammalian milk osteopontin for enhancing immune responsiveness |
KR20230037067A (ko) | 2016-07-06 | 2023-03-15 | 빌딩 블록 뉴트리셔널즈, 엘엘씨 | 영양 조제식 |
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EP1667537A1 (en) | 2006-06-14 |
PL1667537T3 (pl) | 2014-04-30 |
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EP1667537B1 (en) | 2013-11-06 |
AU2004271686B2 (en) | 2009-10-01 |
CA2538836A1 (en) | 2005-03-24 |
KR20060125706A (ko) | 2006-12-06 |
KR101499663B1 (ko) | 2015-03-06 |
CA2538836C (en) | 2013-11-19 |
BRPI0414496B1 (pt) | 2014-05-13 |
BRPI0414496A (pt) | 2006-11-14 |
WO2005025333A1 (en) | 2005-03-24 |
AU2004271686A1 (en) | 2005-03-24 |
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