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  • fractions  (1)
  • protein refolding  (1)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of assisted reproduction and genetics 4 (1987), S. 153-158 
    ISSN: 1573-7330
    Keywords: serum ; fractions ; effects ; in vitro fertilization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To examine the effect of various fractions of human fetal cord serum (HCS) on mouse embryos cultured in vitro, heat-inactivated HCS was separated by ultrafiltration into five distinct fractions: Fractions A, MW〉30,000; B, MW 30,000−10,000; C, MW 10,000−5000; D, MW 5000−1000; and E, MW 〈1000. Seven hundred twentyeight single-cell embryos were cultured in TYH- 280 medium supplemented with 8 mg/ml bovine serum albumin (BSA) and a 20% concentration of Fraction A, B, C, D, or E, whole HCS, or BSA alone. Embryos cultured with Fraction A or E or whole HCS demonstrated a significantly reduced growth rate (P〈0.01), while embryos cultured with Fraction D demonstrated a significantly increased growth rate (P〈0.01). Additionally, 649 singlecell embryos were cultured in medium which was supplemented with 8 mg BSA/ml and a 0, 1,2, or 5% concentration of Fraction A or E. Fraction E displayed toxicity even at a 1% concentration (P〈 0.07), while Fraction A demonstrated growth inhibition at a 5% concentration (P 〈0.05) but increased the hatching rate at a 1% concentration (P 〈 0.01). Finally, 635 single-cell embryos were cultured with four distinct fractions of HCS obtained from a Sephacryl S-200 column: Fractions I, MW 100,000; II, MW 70,000−100,000; III, MW 30,000−70,000; and IV, low molecular weight (〈5000). Fraction I or III significantly reduced the embryo growth rate as seen with Fraction A (P〈0.01) and Fraction II significantly increased only the hatching rate (P〈0.01), while Fraction IV significantly increased the growth rate as seen with Fraction D. In conclusion, HCS contains embryo growth inhibitory properties in the high (〉30,000) and low (〈1000) molecular weight components, while growth promoting factors are found in the 1000−5000 MW fraction. It also seems that there are some factors in the 70,000−100,000 MW fraction which may promote the ability of the embryo to hatch.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 57 (1998), S. 620-623 
    ISSN: 0006-3592
    Keywords: protein refolding ; reversed micelles ; solid-liquid extraction ; RNase A ; DNA ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This article reports that a reversed micellar solution is useful for refolding proteins directly from a solid source. The solubilization of denatured RNase A, which had been prepared by reprecipitation from the denaturant protein solution, into reversed micelles formulated with sodium di-2-ethylhexyl sulfosuccinate (AOT) has been investigated by a solid-liquid extraction system. This method is an alternative to the ordinary protein extraction in reversed micelles based on the liquid-liquid extraction. The solid-liquid extraction method was found to facilitate the solubilization of denatured proteins more efficiently in the reversed micellar media than the ordinary phase transfer method of liquid extraction. The refolding of denatured RNase A entrapped in reversed micelles was attained by adding a redox reagent (reduced and oxidized glutathion). Enzymatic activity of RNase A was gradually recovered with time in the reversed micelles. The denatured RNase A was completely refolded within 30 h. In addition, the efficiency of protein refolding was enhanced when reversed micelles were applied to denatured RNase A containing a higher protein concentration that, in the case of aqueous media, would lead to protein aggregation. The solid-liquid extraction technique using reversed micelles affords better scale-up advantages in the direct refolding process of insoluble protein aggregates. ©1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 620-623, 1998.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
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