Introduction介紹
The integrity of biologic products depends heavily on maintaining a consistent, controlled cold chain during storage and transport. However, freezing, cold storage, and thawing impose considerable stress on these sensitive products, potentially rendering them not only inactive, but also harmful or even life threatening to patients.¹
生物制品的完整性高度依賴于在儲存和運輸過程中維持穩(wěn)定、可控的冷鏈。然而，冷凍、冷藏和解凍會對這些敏感產(chǎn)品造成相當(dāng)大的壓力，不僅可能導(dǎo)致其失效，還可能對患者造成傷害甚至危及生命。
Therefore, robust cold chain management is essential to preserving product quality, particularly during the drug substance transfer to drug product facility. Furthermore, effective cold chain management is key to maximizing product quality, identity, purity and potency. The challenge lies in the complexity of the process: multiple steps and stakeholders are involved in the packaging, freezing, transport, storage, and thawing of biologics.
因此，穩(wěn)健的冷鏈管理對于保持產(chǎn)品質(zhì)量至關(guān)重要，尤其是在原料藥轉(zhuǎn)移到制劑生產(chǎn)設(shè)施的過程中。此外，有效的冷鏈管理是提高產(chǎn)品質(zhì)量、鑒別、純度和效力的關(guān)鍵。挑戰(zhàn)在于該過程的復(fù)雜性：生物制品的包裝、冷凍、運輸、儲存和解凍涉及多個步驟和多個利益相關(guān)部門。
From upstream and downstream bioprocessing, through to final fill/finish  into  drug  product  containers. The biomanufacturing process presents numerous technical challenges that must be addressed to ensure drugs perform with the highest safety,  efficacy and tolerability in  patients. More than that, each biopharmaceutical product exhibits unique characteristics. Particularly during freezing and thawing, inadequate control of cooling and warming rates can adversely affect product integrity.²
從上游和下游生物加工，到最終灌裝/包裝到制劑容器中，生物制藥工藝面臨著諸多技術(shù)挑戰(zhàn)，必須加以解決，以確保藥物在患者身上發(fā)揮的安全性、有效性和耐受性。此外，每種生物制藥產(chǎn)品都具有獨特的特性。尤其是在冷凍和解凍過程中，冷卻和升溫速率控制不當(dāng)會對產(chǎn)品完整性產(chǎn)生不利影響。
Innovations in cold chain technologies and logistics are therefore essential to support  progress in the biopharmaceutical manufacturing process. These include robust handling strategies for biologics contained in single-use bags and bottles as their primary packaging across the biomanufacturing cold chain, including freezing, refrigerated or frozen storage, cold shipment, and thawing.
因此，冷鏈技術(shù)和物流方面的創(chuàng)新對于支持生物制藥生產(chǎn)工藝的進(jìn)步至關(guān)重要。這些創(chuàng)新包括針對以一次性袋子和瓶子作為主要包裝的生物制劑，制定貫穿整個生物制藥冷鏈（包括冷凍、冷藏或冷凍儲存、冷鏈運輸和解凍）的穩(wěn)健處理策略。

Figure 1: Flow diagram showing cold chain management and distribution steps for biologics contained in single-use bags and bottles
圖 1：流程圖，展示了裝在一次性袋和瓶中的生物制品的冷鏈管理和配送步驟。

Manufacturing challenges 制造挑戰(zhàn)
Ensuring product quality 確保產(chǎn)品質(zhì)量
Biologics are susceptible to physical and chemical degradation. Even slight temperature fluctuations can lead to protein denaturation or aggregation, reducing efficacy. Freeze-thaw cycles of biologics in large-volume formats are often required for their storage and transport; however this process can compromise product integrity. To mitigate these risks, manufacturers must rely on validated processes and advanced monitoring systems that maintain consistent conditions throughout the freezing and thawing process.
生物制劑易受物理和化學(xué)因素的影響而降解。即使是輕微的溫度波動也可能導(dǎo)致蛋白質(zhì)變性或聚集，從而降低療效。大體積生物制劑的儲存和運輸通常需要反復(fù)凍融；然而，這一過程可能會損害產(chǎn)品的完整性。為了降低這些風(fēng)險，生產(chǎn)商必須依賴經(jīng)過驗證的工藝和監(jiān)控系統(tǒng)，以確保在整個凍融過程中保持條件的一致性。

Selecting primary packaging 選擇初級包裝
Single-use containers, primarily bags or bottles, are the most common types of containers  used to transport and store biologics, but each type of container has its own limitations.  Bags offer flexibility and facilitate aseptically closed systems. However, they are prone  to  damage due to mechanical stress and can break if not adequately protected when in a frozen state. Rigid containers also tend to be bulky, which means they take a long time to freeze and thaw. This can compromise product quality. Bottles provide better structural integrity but can limit scalability and require specialized handling equipment. The choice of primary  packaging used during freezing often determines the subsequent cold chain management  technologies employed. This is because not all cold chain management solutions are  compatible with all types of primary packaging.
一次性容器，主要是瓶子或袋子，是運輸和儲存生物制劑容器類型，但每種容器都有其自身的局限性。袋子容器具有靈活性，便于建立無菌封閉系統(tǒng)。然而，它們?nèi)菀滓驒C(jī)械應(yīng)力而損壞，如果在冷凍狀態(tài)下保護(hù)不當(dāng)，則可能破裂。硬質(zhì)容器通常體積較大，這意味著它們需要較長的凍融時間。這可能會影響產(chǎn)品質(zhì)量。瓶裝容器具有更好的結(jié)構(gòu)完整性，但會限制其規(guī)?；a(chǎn)，并且需要專門的操作設(shè)備。冷凍過程中使用的初級包裝的選擇通常決定了后續(xù)冷鏈管理技術(shù)的應(yīng)用。這是因為并非所有冷鏈管理解決方案都與所有類型的初級包裝兼容。

Incompatible processes 不兼容的工藝
The use of single-use bags can constrain process flexibility because of equipment incompatibilities and vendor lock-in. Therefore, scalable biopharmaceutical manufacturing solutions must support variable batch volumes while minimizing the need for substantial facility reconfiguration. Bag-independent and modular solutions are thus increasingly popular. Combining modular systems with automation can streamline workflows, reduce the need for manual intervention, and minimize contamination risks, as recommended in EU GMP Annex 1. Ultimately, an end-to-end process that is compatible with every step of cold chain management – from filling and freezing to storage and thawing – ensures a smooth  transition, while also facilitating compliance with relevant quality control standards, such as EU GMP Annex 1.
由于設(shè)備不兼容和供應(yīng)商鎖定，使用一次性袋子會限制工藝的靈活性。因此，可擴(kuò)展的生物制藥生產(chǎn)解決方案必須支持不同批次量，同時減少對設(shè)施進(jìn)行大規(guī)模重新配置的需求。因此，獨立于袋子的模塊化解決方案越來越受歡迎。將模塊化系統(tǒng)與自動化相結(jié)合可以簡化工作流程，減少人工干預(yù)，并降低污染風(fēng)險，正如歐盟GMP附錄1所建議的那樣。最終，一個與冷鏈管理的每個步驟（從灌裝和冷凍到儲存和解凍）都兼容的端到端流程，可以確保平穩(wěn)過渡，同時也有助于符合相關(guān)的質(zhì)量控制標(biāo)準(zhǔn)，例如歐盟GMP附錄1。

Sustainable processes 可持續(xù)過程
Cold chain operations are energy intensive, driven largely by the power requirements of freezers and temperature-controlled storage units. Optimizing freezing profiles and investing in energy-efficient equipment can help reduce the environmental impact of these operations. The use of fluorinated gases, also known as F-gases, is gradually being phased out, with the European Union setting a goal for a complete phase-out by 2050.³ They are being replaced by natural gases and air-cooled technologies with much lower global warming potential (GWP) than F-gases.
冷鏈操作是能源密集型的，這主要是由于冷凍機(jī)和溫控存儲單元的電力需求造成的。優(yōu)化冷凍工藝流程并投資節(jié)能設(shè)備有助于減少這些作業(yè)對環(huán)境的影響。含氟氣體（也稱氟化氣體）的使用正在逐步淘汰，歐盟的目標(biāo)是在2050年前淘汰。3取而代之的是天然氣和空氣冷卻技術(shù)，這些技術(shù)的變暖潛值（GWP）遠(yuǎn)低于氟化氣體。

Figure 2: Reductions in product quality during cold chain management processes are a major concern for biopharmaceutical manufacturers
圖 2：冷鏈管理過程中產(chǎn)品質(zhì)量下降是生物制藥生產(chǎn)商關(guān)注的主要問題。


The behavior of biologics during freezing
生物制品在冷凍過程中的行為
When biologics are frozen, several important changes occur that can influence their stability and efficacy. A thorough understanding of these behaviors is essential for preserving long-term product quality and efficacy.
生物制劑冷凍保存期間會發(fā)生一些重要變化，這些變化會影響其穩(wěn)定性和療效。深入了解這些變化對于保持產(chǎn)品的長期質(zhì)量和療效至關(guān)重要。

What happens when freezing biologics? 生物制劑冷凍后會發(fā)生什么？
Proteins are thermally unstable molecules, and their native structure depends on a delicate balance of interactions within the protein itself and between the protein and surrounding components, such as solvents. When a protein solution is frozen, the temperature of the solution initially decreases steadily as the heat is removed. As the temperature continues to decrease, the solution can become super-cooled, which means that it drops below its normal freezing point without immediately forming ice. However, at a certain temperature, ice begins to form; this is known as ice nucleation. Once ice starts to form, the temperature briefly stabilizes because energy is being used to change the liquid protein solution into solid ice, not to lower the temperature further. This stage is known as the phase transition. After most of the liquid has turned to ice, the temperature begins to decrease again as more heat is removed, and the solution becomes completely solid.
蛋白質(zhì)是熱穩(wěn)定性較差的分子，其天然結(jié)構(gòu)依賴于蛋白質(zhì)內(nèi)部以及蛋白質(zhì)與周圍成分（例如溶劑）之間相互作用的微妙平衡。當(dāng)?shù)鞍踪|(zhì)溶液被冷凍時，隨著熱量的移除，溶液的溫度最初會穩(wěn)定下降。隨著溫度持續(xù)下降，溶液會變得過冷，這意味著它的溫度會降至正常冰點以下，但不會立即形成冰。然而，在達(dá)到一定溫度后，冰開始形成；這被稱為冰核形成。一旦冰開始形成，溫度會短暫穩(wěn)定下來，因為能量被用于將液態(tài)蛋白質(zhì)溶液轉(zhuǎn)化為固態(tài)冰，而不是用于進(jìn)一步降低溫度。這個階段被稱為相變。當(dāng)大部分液體轉(zhuǎn)化為冰后，隨著更多熱量的移除，溫度再次開始下降，溶液最終凝固。

Figure 3: The behavior of biologics during the freezing process 圖 3：生物制品在冷凍過程中的行為


Figure 4: Ice crystallization at each step in the freezing process

What physical changes impact protein stability during freezing?
冷凍過程中哪些物理變化會影響蛋白質(zhì)的穩(wěn)定性？
Freezing and thawing can alter how proteins interact with their environment, which in turn can reduce their stability. One of the most important effects is cryoconcentration.^4,5 When pharmaceutical products are frozen in large batches, ice forms as the temperature decreases;  this removes water that normally hydrates and stabilizes proteins. As a result, proteins and  other formulation components become concentrated in the remaining unfrozen solution. The loss of protective water and the increased concentration of other ingredients can destabilize proteins, leading to aggregation, partial unfolding, or unwanted interactions with formulation components or container surfaces.
凍融過程會改變蛋白質(zhì)與其環(huán)境的相互作用方式，進(jìn)而降低其穩(wěn)定性。其中最重要的影響之一是冷凍濃縮。當(dāng)藥品批量冷凍時，隨著溫度降低會形成冰；冰會帶走通常用于水合和穩(wěn)定蛋白質(zhì)的水分。因此，蛋白質(zhì)和其他制劑成分會在剩余的未凍結(jié)溶液中濃縮。保護(hù)性水分的損失和其他成分濃度的增加會導(dǎo)致蛋白質(zhì)不穩(wěn)定，進(jìn)而引起聚集、部分展開或與制劑成分或容器表面發(fā)生不必要的相互作用。
Cryoconcentration can introduce several additional destabilizing effects. As freezing progresses, the solution becomes more viscous, and the rates of freezing and thawing influence how long these altered conditions persist. Ice formation can also trap small air bubbles, which may further disrupt protein structure and reduce stability. Another key factor is the formation of ice-liquid interfaces.⁵ Proteins that adsorb to these interfaces can undergo structural changes or denaturation; therefore, control of the cooling rate is critical. More rapid freezing generally produces smaller ice crystal s and results in a larger ice-liquid interface. This can help limit the unwanted crystallization of buffering components and support  maintenance of solution pH.
冷凍濃縮還會引入其他幾種不穩(wěn)定因素。隨著冷凍的進(jìn)行，溶液粘度會增加，而凍融速率會影響這些改變狀態(tài)的持續(xù)時間。冰的形成還會捕獲微小的氣泡，這可能會進(jìn)一步破壞蛋白質(zhì)結(jié)構(gòu)并降低其穩(wěn)定性。另一個關(guān)鍵因素是冰-液界面的形成。吸附在這些界面上的蛋白質(zhì)可能會發(fā)生結(jié)構(gòu)變化或變性；因此，控制冷卻速率至關(guān)重要?？焖倮鋬鐾ǔa(chǎn)生更小的冰晶，并形成更大的冰液界面。這有助于限制緩沖成分的非預(yù)期結(jié)晶，并有助于維持溶液的pH值。

What does this mean when freezing biologics during manufacturing?
在生產(chǎn)過程中冷凍生物制劑意味著什么？
Ultimately, freezing is not just a matter of lowering the temperature. For example, fast freezing is optimal for most proteins, such as monoclonal antibodies, whereas slower freezing rates are preferable for other proteins.⁶
歸根結(jié)底，冷凍不僅僅是降低溫度的問題。例如，快速冷凍對大多數(shù)蛋白質(zhì)（如單克隆抗體）來說是好選擇，而較慢的冷凍速率則更適合其他蛋白質(zhì)。
Precise control over process parameters is required to minimize destabilizing effects and preserve the integrity of biologics. Manufacturers must account for these physical changes and their impact on protein stability to ensure consistent product quality and reliable handling of bulk drug substances.
精確控制工藝參數(shù)對于減少不穩(wěn)定因素并保持生物制劑的完整性至關(guān)重要。生產(chǎn)商必須考慮這些物理變化及其對蛋白質(zhì)穩(wěn)定性的影響，以確保產(chǎn)品質(zhì)量的一致性和原料藥處理的可靠性。

Figure 5: RoSS® shel l is a protective secondary packaging for all two-dimensional single-use bags currently on the market
圖 5：RoSS® shell 是一種適用于目前市面上所有一次性2D袋子的保護(hù)性二級包裝。


Best practice: Controlled freezing of biologics
The freezing process has profound implications for the stability of protein-based biologics. Cryoconcentration and the formation of ice-liquid interfaces can lead to the loss of protein structure and function if not properly controlled. The extent of these effects depends on a variety of factors, including:
冷凍過程對蛋白質(zhì)生物制劑的穩(wěn)定性有著深遠(yuǎn)的影響。如果控制不當(dāng)，冷凍濃縮和冰液界面的形成會導(dǎo)致蛋白質(zhì)結(jié)構(gòu)和功能的喪失。這些影響的程度取決于多種因素，包括：

• Freezing and thawing rates 冷凍和解凍速率

• Single-use container geometry 一次性容器幾何形狀

• Processed volume 處理體積

Therefore, effective management of these physical changes requires controlled freezing  strategies for different container types, such as single-use bags and bottles, and for a range of processing volumes.
因此，要有效管理這些物理變化，需要針對不同類型的容器（如一次性袋子和瓶子）以及不同的加工量制定受控的冷凍策略。

Freezing biologics in single-use bags 將生物制劑冷凍于一次性袋子中
2D bioprocess containers that have a small water column can be frozen using plate-freezing technology, which enables more rapid, controlled freezing than traditional static freezing. When employing plate-freezing technology with single-use bags, the bags should be placed in specialized secondary packaging designed to ensure optimal heat transfer. Rapid, controlled freezing using plate freezers enables simultaneous freezing of the drug substance in single-use bags from both the top and the bottom. This helps prevent cryoconcentration of  the drug  substances and other formulation components in the center of the bag. Both conductive  cooling, which provides direct contact with the cooling medium as in plate freezers, and convective cooling, which uses an airflow-based technology as in blast freezers, achieve faster and more homogeneous freezing than static freezers. Figure 6 illustrates the difference in freezing results between static freezers and plate freezers. The degree of cryoconcentration in a single-use bag compared to the initial liquid-state concentration when frozen in a static freezer is significantly higher than that observed when plate-freezing is used. A study conducted in collaboration with Zurich University of Applied Sciences measured cryoconcentration levels of up to +212 % after freezing in a static freezer, whereas cryoconcentration levels using plate freezers showed no major changes.⁷
對于含有少量水柱的二維生物工藝容器，可采用平板冷凍技術(shù)進(jìn)行冷凍，該技術(shù)比傳統(tǒng)的靜態(tài)冷凍技術(shù)能夠?qū)崿F(xiàn)更快速、更可控的冷凍。當(dāng)使用平板冷凍技術(shù)處理一次性袋子時，應(yīng)將袋子置于專門設(shè)計的二級包裝中，以確保最佳的熱傳遞。平板冷凍機(jī)能夠從頂部和底部同時冷凍一次性袋子中的藥物，從而實現(xiàn)快速、可控的冷凍。這有助于防止藥物和其他制劑成分在袋子中心發(fā)生冷凍濃縮。無論是傳導(dǎo)冷卻（如平板冷凍機(jī)中與冷卻介質(zhì)直接接觸）還是對流冷卻（如速凍機(jī)中利用氣流技術(shù)），都能比靜態(tài)冷凍機(jī)實現(xiàn)更快、更均勻的冷凍。圖 6 展示了靜態(tài)冷凍機(jī)和平板冷凍機(jī)冷凍結(jié)果的差異。與靜態(tài)冷凍機(jī)相比，使用平板冷凍技術(shù)冷凍的一次性袋子中的冷凍濃縮程度明顯更高。與蘇黎世應(yīng)用科學(xué)大學(xué)合作進(jìn)行的一項研究測量了靜態(tài)冷凍機(jī)冷凍后的冷凍濃縮水平高達(dá) +212 %，而使用平板冷凍機(jī)的冷凍濃縮水平則沒有出現(xiàn)重大變化。

Figure 6: Visualized cryoconcentration of uncontrolled conventional freezing (top) vs controlled plate-freezing (bottom)
圖 6：非受控常規(guī)冷凍（上圖）與受控平板冷凍（下圖）的冷凍濃縮可視化圖


Dark blue spots indicate greater cryoconcentration. Rapid and controlled freezing with a plate freezer enables more homogeneous freezing than conventional freezing.
深藍(lán)色斑點表示冷凍濃縮程度更高。與傳統(tǒng)冷凍方法相比，使用平板冷凍機(jī)進(jìn)行快速可控冷凍能夠?qū)崿F(xiàn)更均勻的冷凍效果。
As noted earlier, small air bubbles can occur during the cooling process, which can further affect proteins and reduce their stability. Stress at the liquid-ice interface during the formation of ice can also destabilize proteins.8 So how can these effects be prevented to achieve a homogeneous freezing result? Single Use Support has performed computed tomography (CT) scans to visualize when these phenomena occur to identify ways in which they can be avoided. In Figure 7, an uneven freezing result following static freezing is clearly visible. Bubbles and cracks at the final point of solidification are visible, indicating cryoconcentration.
如前所述，冷卻過程中可能會產(chǎn)生微小氣泡，這會進(jìn)一步影響蛋白質(zhì)并降低其穩(wěn)定性。冰形成過程中液態(tài)-冰界面處的應(yīng)力也會破壞蛋白質(zhì)的穩(wěn)定性。8 那么，如何避免這些影響以獲得均勻的冷凍效果呢？Single Use Support 公司進(jìn)行了計算機(jī)斷層掃描 (CT)，以觀察這些現(xiàn)象發(fā)生的時間，并找出避免它們的方法。圖 7 清晰地顯示了靜態(tài)冷凍后冷凍效果不均勻的情況。在最終凝固點處可以看到氣泡和裂紋，表明存在低溫濃縮現(xiàn)象。
Figure 7: CT scan cross-sectional views (horizontal and vertical) of a single-use bag frozen in a static freezer show uneven freezing, with bubbles and cracks concentrated at the final point of solidification
圖 7：在靜態(tài)冰箱中冷凍的一次性袋子的 CT 掃描橫截面視圖（水平和垂直）顯示冷凍不均勻，氣泡和裂紋集中在最終凝固點。

Figure 8: CT scan cross-sectional views (horizontal and vertical) of a single-use bag frozen using a plate freezer show homogeneous ice fronts with no bubbles or cracks
圖 8：使用平板冷凍機(jī)冷凍的一次性袋子的 CT 掃描橫截面視圖（水平和垂直）顯示均勻的冰層前沿，沒有氣泡或裂紋。


In contrast, Figure 8 shows homogeneous freezing, achieved by following freezing protocols to obtain optimal freezing rates and gradients, using Single Use Support’s plate freezer, RoSS. pFTU. This fully automated, protocol-driven approach not only aligns with US Food and Drug Administration 21 CFR Part 11 standards for electronic records and delivers consistent results across a variety of scales but also maximizes product integrity. The controlled-rate plate freezer RoSS.pFTU, used in combination with Single Use Support’s protective secondary packaging RoSS^® shell, can achieve maximum product quality during the freezing of biologics.
相比之下，圖8展示了使用Single Use Support公司的 RoSS.pFTU平板冷凍機(jī)，通過遵循冷凍方案獲得冷凍速率和梯度，從而實現(xiàn)均勻冷凍。這種全自動、配方驅(qū)動的方法不僅符合 (FDA) 21 CFR Part 11 的電子記錄標(biāo)準(zhǔn)，并且在各種規(guī)模下都能提供一致的結(jié)果，而且還能保證產(chǎn)品的完整性。RoSS.pFTU 可控速率平板冷凍機(jī)與 Single Use Support 公司的 RoSS^® 保護(hù)性二級包裝外殼配合使用，可在生物制品冷凍過程中實現(xiàn)產(chǎn)品質(zhì)量。
The RoSS® shell offers robust, reliable, and secure protection for single-use bioprocess containers of any size and vendor during freezing, transportation, storage, and thawing.
RoSS® 外殼為任何尺寸和供應(yīng)商的一次性生物工藝容器在冷凍、運輸、儲存和解凍過程中提供*、可靠和安全的保護(hù)。
With its innovative design, the RoSS^® shell is a tamper-evident system that minimizes contamination risk and ensures all single-use bags are efficiently cooled using plate freezing technology while providing protection during storage and shipping. This:
憑借其創(chuàng)新設(shè)計，RoSS® 外殼是一個防篡改系統(tǒng)，可降低污染風(fēng)險，并確保所有一次性袋子均能使用平板冷凍技術(shù)進(jìn)行高效冷卻，同時在儲存和運輸過程中提供保護(hù)。

• Minimizes product losses attributable to bag breakages

• 減少因袋子破損造成的產(chǎn)品損失

• Maximizes product quality by providing optimal freezing and thawing conditions

• 通過提供冷凍和解凍條件，提高產(chǎn)品質(zhì)量

• Is compatible with all types of two-dimensional (2D) single-use bags

• 兼容所有類型的二維（2D）一次性袋子

Single-use bags secured within the robust RoSS^® shell can be placed into the RoSS.pFTU plate-based freezer, which:
一次性袋子固定在堅固的 RoSS® 外殼內(nèi)，可放入 RoSS.pFTU 平板式冷凍機(jī)中，該冷凍機(jī)：

• Enables controlled freezing down to -80 °C with the highest possible speed and accuracy

• 能夠以高速度和精度實現(xiàn)低至-80°C的受控冷凍

• Offers a capacity of up to 400 L, depending on bag type

• 容量高達(dá)400升，具體取決于袋子類型

• Is fully compatible with 2D single-use bags of any size and from any vendor

• 兼容任何尺寸、任何供應(yīng)商的二維一次性袋子

• Can achieve optimal product stability results for bulk drug substances, including monoclonal antibodies

• 可實現(xiàn)原料藥（包括單克隆抗體）的產(chǎn)品穩(wěn)定性

• Is fully automated and ready for cGMP use

• 全自動運行，符合cGMP要求

Figure 9: Fully loaded RoSS.pFTU Large Scale
圖 9：滿載的 RoSS.pFTU 大規(guī)模  


Freezing biologics in bottles 將生物制劑冷凍于瓶中
Unlike single-use bags, bottles present unique challenges when used to freeze biologics; this is due to their bulk and their requirement for vertical placement during freezing. It is therefore recommended to freeze bottles using static or blast freezing methods.
與一次性袋子不同，瓶子在冷凍生物制劑時面臨著獨特的挑戰(zhàn)；這是由于瓶子體積較大，且冷凍過程中需要垂直放置。因此，建議采用靜態(tài)冷凍或速凍方法冷凍瓶子。
However, uncontrolled freezing in bottles often leads to uneven temperature gradients and cryoconcentration, compromising product quality. Blast freezing, particularly with forced air flow, is superior to static freezing methods but still requires careful control.
然而，瓶內(nèi)冷凍若不受控制，往往會導(dǎo)致溫度梯度不均勻和濃縮，從而影響產(chǎn)品質(zhì)量。速凍，特別是采用強(qiáng)制氣流的速凍，優(yōu)于靜態(tài)冷凍方法，但仍需嚴(yán)格控制。
Studies have shown that uncontrolled freezing can result in considerable differences in solute concentration between the top and bottom of a bottle, indicating heterogeneous freezing and reduced reproducibility, with cryoconcentration levels increasing by more than 300 %.⁹
研究表明，冷凍若不受控制，會導(dǎo)致瓶內(nèi)頂部和底部溶質(zhì)濃度出現(xiàn)顯著差異，表明冷凍不均勻，重復(fù)性降低，濃縮程度增加超過300%。
As a bottle is subjected to the freezing process, pressure on the side walls from the growing ice fronts pushes liquid toward the center, forming a cone at the top. This phenomenon is known as the “volcano effect”, illustrated in Figure 10.
當(dāng)瓶子處于冷凍過程中時，不斷增長的冰鋒對瓶壁施加壓力，將液體推向中心，在頂部形成錐形。這種現(xiàn)象被稱為“火山效應(yīng)”，如圖10所示。

Figure 10: Visualized cryoconcentration in bottles under uncontrolled freezing conditions showing the “volcano effect” (left) compared with controlled blast freezing (right)
圖 10：在不受控制的冷凍條件下，瓶中低溫濃縮的可視化結(jié)果，顯示了“火山效應(yīng)”（左圖），并與受控的速凍（右圖）進(jìn)行了比較。

To address these issues, Single Use Support has developed controlled-rate freezing protocols for bottles. For large bottles, freezing rates of approximately -0.1 °C to -0.5 °C/min are essential for maintaining protein stability.4,10 Controlled air distribution with advanced air flow systems optimizes the freezing behavior of drug substances to ensure bottle-to-bottle reproducibility and homogeneous freezing results. Visualization of heat transfer and airflow patterns demonstrates that high-performance forced air distribution enables consistent temperature control and uniform freezing. Single Use Support’s RoSS.BLST is a blast freezer that controls the rate of cooling by employing smart air distribution, as shown in Figure 11. It is a modular, controlled-rate blast freezer that incorporates forced-air  technology. Its best-in-class chamber size-to-foot-print ratio enables manufacturers to efficiently freeze bulk-packaged biologics. It can accommodate bottles, bulky single-use bag setups, and other single-use containers. For example, it can accommodate up to 192 2 L bottles and can freeze drug substances according to optimal freezing protocols to achieve maximum product stability. It also contributes to efficient and sustainable freezing  by  providing great process flexibility:
為了解決這些問題，Single Use Support 開發(fā)了適用于瓶裝產(chǎn)品的控溫速凍方案。對于大瓶裝產(chǎn)品，約 -0.1℃ 至 -0.5 °C/min 的速凍速率對于維持蛋白質(zhì)穩(wěn)定性至關(guān)重要^4,10。采用氣流系統(tǒng)的可控空氣分配能夠優(yōu)化藥物的冷凍行為，確保瓶與瓶之間冷凍效果的可重復(fù)性和均勻性。熱傳遞和氣流模式的可視化結(jié)果表明，高性能強(qiáng)制空氣分配能夠?qū)崿F(xiàn)穩(wěn)定的溫度控制和均勻的冷凍效果。如圖 11 所示，Single Use Support 的 RoSS.BLST 是一款采用智能空氣分配控制冷卻速率的速凍機(jī)。它是一款模塊化、控溫速凍機(jī)，并采用了強(qiáng)制空氣技術(shù)。其腔室尺寸與占地面積比使生產(chǎn)商能夠高效地冷凍散裝生物制劑。它可以容納瓶裝產(chǎn)品、體積較大的一次性袋子以及其他一次性容器。例如，它最多可容納 192 瓶 2 升裝藥品，并可根據(jù)冷凍方案冷凍藥品，以實現(xiàn)產(chǎn)品穩(wěn)定性。此外，它還具有高的工藝靈活性，有助于實現(xiàn)高效且可持續(xù)的冷凍：

• Modular shelving for bottle sizes ranging from 30 mL to 10 L

• 模塊化貨架，適用于30毫升至10升的瓶裝飲料

• Sustainable design with natural refrigerants and a low refrigerant load

• 采用可持續(xù)設(shè)計，使用天然制冷劑，制冷劑負(fù)荷低

• Complies with EN 378 and ISO 5149 standards

• 符合EN 378和ISO 5149標(biāo)準(zhǔn)

• Includes thawing functionality with integrated shaking

• 內(nèi)置搖晃功能，可解凍飲料

Figure 11: Visualization of heat transfer and airflow patterns when using RoSS.BLST for temperature control , showing the high-performance forced air distribution when used with single-use bottles (top) and bags (bottom)
圖 11：使用 RoSS.BLST 進(jìn)行溫度控制時的熱傳遞和氣流模式可視化，顯示了與一次性瓶子（上圖）和袋子（下圖）一起使用時的高性能強(qiáng)制空氣分配。


Bottle RoSS protects bottle caps with attached tubing assemblies. It provides tamper-evident integrity and reduces product losses during cold chain storage and shipping. Therefore, Bottle RoSS ensures the safe, efficient, and scalable management of biologics in bottles, regardless of bottle manufacturer or type.
Bottle RoSS 通過連接的管路組件保護(hù)瓶蓋。它提供防篡改保護(hù)，并減少冷鏈儲存和運輸過程中的產(chǎn)品損失。因此，無論瓶子制造商或類型如何，Bottle RoSS 都能確保對瓶裝生物制劑進(jìn)行安全、高效且可擴(kuò)展的管理。

Figure 12: A RoSS.BLST fully loaded with 2 L bottles 圖 12：裝滿 2 升瓶子的 RoSS.BLST

Figure 13: Bottle RoSS for protection of single-use assemblies on bottles during cold chain handling
圖 13：瓶身 RoSS，用于在冷鏈搬運過程中保護(hù)瓶子上的一次性組件。


Maintaining cold chain for storage and transport
維持冷鏈以進(jìn)行儲存和運輸
The efficient and safe freezing of biologics in single-use bags and bottles requires a fully closed, Annex 1-aligned, and aseptic system during the entire fluid and cold chain management process. The process begins with automated filling and filtration, continues through freezing, cold storage and shipping, and concludes with controlled thawing at the drug product site, until the drug substance is removed from the singleuse containers for fill/finish. The integration of closed systems at every stage minimizes contamination risks, streamlines workflows, and ensures product integrity. This end-toend strategy fosters compatibility between single-use technologies, regardless of the type or size of primary packaging used by the manufacturer. Furthermore, closed systems fulfill GMP-relevant standards, and can be seamlessly integrated into customer networks for process control. An automated and compatible cold chain improves operational efficiency, making it the gold standard for the management of single-use containers used in biopharmaceutical manufacturing, such as single-use bags and bottles.
生物制劑在一次性袋子和瓶中的高效安全冷凍，需要在整個液體和冷鏈管理過程中采用封閉、符合附件1要求且無菌的系統(tǒng)。該流程始于自動化灌裝和過濾，貫穿冷凍、冷藏和運輸，最終在藥品生產(chǎn)現(xiàn)場進(jìn)行受控解凍，直至將原料藥從一次性容器中取出進(jìn)行灌裝/包裝。在每個階段集成封閉系統(tǒng)可限度地降低污染風(fēng)險，簡化工作流程，并確保產(chǎn)品完整性。這種端到端策略促進(jìn)了各種一次性技術(shù)之間的兼容性，無論制造商使用何種類型或尺寸的初級包裝。此外，封閉系統(tǒng)符合GMP相關(guān)標(biāo)準(zhǔn)，并且可以無縫集成到客戶網(wǎng)絡(luò)中進(jìn)行過程控制。自動化且兼容的冷鏈提高了運營效率，使其成為生物制藥生產(chǎn)中一次性容器（例如一次性袋子和瓶）管理的黃金標(biāo)準(zhǔn)。

Cold storage of biologics 生物制劑的冷藏
For ultra-low temperature storage, the RoSS.ULTF platform provides secure, scalable cold storage down to -80 °C. The modul ar interior can accommodate either single-use bags protected in RoSS® shells or single-use bottles and their assemblies protected with Bottle RoSS, as well as other bulk-packaged drug substances of various sizes and batch volumes. This design ensures compatibility with all container types and safety for high-value drug substances. Automated monitoring and GMP-compatible documentation support regulatory requirements and batch traceability.
RoSS.ULTF平臺專為超低溫儲存而設(shè)計，可提供安全、可擴(kuò)展的冷藏環(huán)境，溫度可達(dá)-80°C。其模塊化內(nèi)部結(jié)構(gòu)可容納RoSS®外殼保護(hù)的一次性袋子、RoSS®瓶裝保護(hù)的一次性藥瓶及其組件，以及其他各種規(guī)格和批次的散裝藥品。該設(shè)計確保與所有容器類型兼容，并保障高價值藥品的安全性。自動化監(jiān)控和符合GMP規(guī)范的文檔記錄功能可滿足監(jiān)管要求和批次追溯性。

Cold chain shipping container 冷鏈運輸集裝箱
Maintaining cold chain is critical when transporting bulk drug substances. The RoSS.SHIP solution enables safe, temperature-controlled shipping of bottles and bags. This container is specifically designed to maintain cold chain, ensuring that bulk drug substances remain at temperatures below -60°C for several days during transport. Integrated track and trace technology enables real-time monitoring of temperature and location, providing full visibility and compliance throughout transit. RoSS.SHIP’s robust packaging minimizes the risk of breakage, contamination, and product loss, ensuring that biologics arrive at their destination in optimal condition.
在運輸原料藥時，維持冷鏈至關(guān)重要。RoSS.SHIP解決方案可實現(xiàn)瓶裝和袋裝原料藥的安全溫控運輸。該容器專為維持冷鏈而設(shè)計，確保原料藥在運輸過程中數(shù)日內(nèi)保持在-60°C以下的溫度。集成的追蹤溯源技術(shù)可實時監(jiān)控溫度和位置，從而在整個運輸過程中提供全面的可視性和合規(guī)性。RoSS.SHIP堅固的包裝限度地降低了破損、污染和產(chǎn)品損失的風(fēng)險，確保生物制劑以好狀態(tài)抵達(dá)目的地。

Figure 14: Ultra-cold storage of biologics with RoSS.ULTF
圖 14：采用 RoSS.ULTF 進(jìn)行生物制劑的超低溫儲存

Figure 15: The cold chain shipping container RoSS.SHIP
圖 15：冷鏈運輸集裝箱 RoSS.SHIP

Thawing biologics at the drug product site 在藥品生產(chǎn)現(xiàn)場解凍生物制劑