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    Heat Inactivation of Serum

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    Vicnt Escamilla
    Heat inactivation of serum is necessary to eliminate complement-mediated cell lysis since serum contains complement. Heat inactivation destroys complement but can also damage heat-labile growth factors, vitamins, and hormones in serum. The standard procedure is to heat serum to 56°C for 30 minutes while constantly swirling, as this inactivates complement without excessive damage to other components. Care must be taken to slowly thaw and heat serum under 56.3°C for no more than 30 minutes to prevent protein denaturation and turbidity.

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    Heat Inactivation of Serum

    Uploaded by

    Vicnt Escamilla
    130 views1 page
    Heat inactivation of serum is necessary to eliminate complement-mediated cell lysis since serum contains complement. Heat inactivation destroys complement but can also damage heat-labile growth factors, vitamins, and hormones in serum. The standard procedure is to heat serum to 56°C for 30 minutes while constantly swirling, as this inactivates complement without excessive damage to other components. Care must be taken to slowly thaw and heat serum under 56.3°C for no more than 30 minutes to prevent protein denaturation and turbidity.

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    Heat_Inactivation_of_Serum

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    Heat inactivation of serum is necessary to eliminate complement-mediated cell lysis since serum contains complement. Heat inactivation destroys complement but can also damage heat-labile growth factors, vitamins, and hormones in serum. The standard procedure is to heat serum to 56°C for 30 minutes while constantly swirling, as this inactivates complement without excessive damage to other components. Care must be taken to slowly thaw and heat serum under 56.3°C for no more than 30 minutes to prevent protein denaturation and turbidity.

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    Attribution Non-Commercial (BY-NC)
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    130 views1 page

    Heat Inactivation of Serum

    Uploaded by

    Vicnt Escamilla
    Heat inactivation of serum is necessary to eliminate complement-mediated cell lysis since serum contains complement. Heat inactivation destroys complement but can also damage heat-labile growth factors, vitamins, and hormones in serum. The standard procedure is to heat serum to 56°C for 30 minutes while constantly swirling, as this inactivates complement without excessive damage to other components. Care must be taken to slowly thaw and heat serum under 56.3°C for no more than 30 minutes to prevent protein denaturation and turbidity.

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    Technical Information

    Heat Inactivation of Serum

    Serum is a very complex supplement containing mostly Since serum is a blood product, it contains complement,
    proteins, but also growth factors, hormones, amino acids, which can lead to complement-mediated cell lysis. To elimi-
    glucose, trypsin inhibitors, and lipids. The most common nate this risk, serum can be heat-inactivated. Heat inacti-
    types of sera include: fetal bovine (FBS), newborn calf vation destroys the complement, but can also destroy heat-
    (NCS), horse (equine), pig (porcine), and human. There labile growth factors, vitamins, amino acids, and hormones
    are also several different forms of serum due to the types present in serum. Damage to these from extended heating
    of processing available. Fetal bovine serum (FBS) has been or from temperatures greater than 56C will decrease cell
    adopted as the standard supplement because of its rich growth. Heating oftentimes can cause the proteins and
    content of growth factors and its low gamma globulin con- lipids concentrated at the bottom to appear as white glob-
    tent. ules. Constant swirling during the heat inactivation pro-
    cess will help solve this problem.
    Some of the major functions of serum are to provide ba-
    sic nutrients, hormones and growth factors, attachment and The following is a suggested procedure for proper heat-
    spreading factors, pH buffer, protease inhibitors, binding inactivation of serum.
    proteins carrying hormones, vitamins, minerals, and lipids.
    Other advantages to using serum include protection against
    toxic effects associated with pH change, the presence of
    heavy metals ions, endotoxin, and proteolytic activity.

    Procedure
    1. Allow frozen serum to thaw under refrigerated conditions or at room temperature. Serum can be thawed in a 37C
    water bath, however monitor the time and remove as soon as the serum is thawed.
    2. Adjust the temperature of a water bath to 56C and allow a thermometer to equilibrate to the temperature of the
    water (i.e. overnight). The thermometer should be placed in a bottle of water, similar to the volume of the bottle of
    serum, to prevent damage to the thermometer.
    3. Once the serum has thawed completely, mix the serum by gently swirling the container to ensure that the solution is
    homogenous and transfer to the water bath. Do not submerge the bottles or allow the water level to extend to the
    caps of the bottles, as they may compromise the integrity of the container.
    4. Monitor the temperature in the water bottle using the thermometer to ensure a steady temperature.
    5. During incubation, swirl the contents of both bottles every five minutes to ensure that the contents are evenly
    heated.
    6. Once the bottle of serum has been at 56C for 30 minutes, transfer the bottle of serum to ice to cool and proceed as
    needed. To prevent multiple freeze-thaw cycles, alliquot serum into smaller sterile containers, label with date and lot
    number, and transfer to a freezer (-20C).

    To prevent protein denaturation, which leads to turbidity and the presence of globules in the serum:
    3 Do not increase the speed of thawing using temperatures greater than 25C.
    3 Do not heat inactivate serum that his partially mixed/partially thawed.
    3 Do not heat inactivate serum at a temperature greater than 56.3C.
    3 Do not inactivate serum for greater than 30C.
    3 Be sure to swirl the serum during the heat inactivation process.

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