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Effect of extracellular pH, ammonia,...

Borys, Michael Christopher.

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Effect of extracellular pH, ammonia, and cell aggregation on the specific expression rate and N-linked glycosylation of recombinant mouse-placental lactogen proteins by Chinese hamster ovary (CHO) cells.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Effect of extracellular pH, ammonia, and cell aggregation on the specific expression rate and N-linked glycosylation of recombinant mouse-placental lactogen proteins by Chinese hamster ovary (CHO) cells./
Author:	Borys, Michael Christopher.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 1993,
Description:	253 p.
Notes:	Source: Dissertation Abstracts International, Volume: 55-01, Section: B, page: 1550.
Contained By:	Dissertation Abstracts International55-01B.
Subject:	Chemical engineering. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9415697

Effect of extracellular pH, ammonia, and cell aggregation on the specific expression rate and N-linked glycosylation of recombinant mouse-placental lactogen proteins by Chinese hamster ovary (CHO) cells.
Borys, Michael Christopher.

Effect of extracellular pH, ammonia, and cell aggregation on the specific expression rate and N-linked glycosylation of recombinant mouse-placental lactogen proteins by Chinese hamster ovary (CHO) cells. - Ann Arbor : ProQuest Dissertations & Theses, 1993 - 253 p.

Source: Dissertation Abstracts International, Volume: 55-01, Section: B, page: 1550.

Thesis (Ph.D.)--Northwestern University, 1993.

Experiments were performed using recombinant, anchorage-dependent Chinese hamster ovary (CHO) cells expressing either the glycosylated recombinant protein mouse placental lactogen-I (mPL-I) or the unglycosylated recombinant protein mouse placental lactogen-II (mPL-II). The mPL-I or mPL-II cDNA was co-amplified with DHFR by methotrexate selection in CHO cells.Subjects--Topical Terms:

560457
Chemical engineering.

Effect of extracellular pH, ammonia, and cell aggregation on the specific expression rate and N-linked glycosylation of recombinant mouse-placental lactogen proteins by Chinese hamster ovary (CHO) cells.
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035 $a (MiAaPQ)AAI9415697
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100 1 $a Borys, Michael Christopher. $3 3283263
245 1 0 $a Effect of extracellular pH, ammonia, and cell aggregation on the specific expression rate and N-linked glycosylation of recombinant mouse-placental lactogen proteins by Chinese hamster ovary (CHO) cells.
260 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 1993
300 $a 253 p.
500 $a Source: Dissertation Abstracts International, Volume: 55-01, Section: B, page: 1550.
500 $a Adviser: E. Terry Papoutsakis.
502 $a Thesis (Ph.D.)--Northwestern University, 1993.
520 $a Experiments were performed using recombinant, anchorage-dependent Chinese hamster ovary (CHO) cells expressing either the glycosylated recombinant protein mouse placental lactogen-I (mPL-I) or the unglycosylated recombinant protein mouse placental lactogen-II (mPL-II). The mPL-I or mPL-II cDNA was co-amplified with DHFR by methotrexate selection in CHO cells.
520 $a Glycosylation patterns and specific expression rates of the recombinant protein mPL-I by CHO cells under non-growth conditions in serum-free, protein expression medium varied significantly over the extracellular pH (pH$\sb{\rm e}$) range of 6.1 to 8.7 in T-flask cultures with monolayer cell coverage. The maximum specific mPL-I expression rates occurred between pH$\sb{\rm e}$ 7.6 and 8.0. The pH$\sb{\rm e}$ effect on protein expression was confirmed using a different CHO cell expressing the unglycosylated recombinant protein mPL-II. The effect of pH$\sb{\rm e}$ on both the total protein secretion and on the glucose uptake by the recombinant CHO cells followed a trend similar to that of the pH$\sb{\rm e}$ effect on recombinant protein expression. Decreases in the extent of N-linked glycosylation of mPL-I were observed at low (below 6.9) and high (above 8.2) pH$\sb{\rm e}$ values.
520 $a The N-linked glycosylation of mPL-I by Chinese hamster ovary (CHO) cell T-flask cultures under non-growth conditions was inhibited by increasing levels of ammonium chloride (3 and 9 mM) in the serum-free, protein expression medium. The effect of ammonia on glycosylation was dependent on the pH$\sb{\rm e}$. At high pH$\sb{\rm e}\ (\ge7.6),$ increasing ammonium chloride concentrations resulted in notable decreases in glycosylation levels. However, at pH$\sb{\rm e}$ 7.2 and 6.7, mPL-I glycosylation was increasingly resistant to elevated ammonium chloride concentrations. There is evidence that the pH$\sb{\rm e}$ dependent effect of ammonium chloride is due to the ammonia species (NH$\sb3).$ Data from both the pH$\sb{\rm e}$ and ammonia experiments indicate that mPL-I with decreased glycosylation may have increased ability to stimulate Nb2 node lymphoma cell proliferation.
520 $a There was preliminary evidence that expression of lower molecular sized mPL-I glycoforms by CHO cells increased from a microcarrier culture with cell aggregates with sizes up to 600 $\mu$m. This effect was dependent upon the culture pH. The addition of 1 $\mu$M or 5 $\mu$M retinoic acid to the serum-free, protein expression medium to confluent T-flask cultures under non-growth conditions was found to decrease mPL-I glycosylation and the specific mPL-I expression rates at both pH$\sb{\rm e}$ 6.7 and 7.6.
590 $a School code: 0163.
650 4 $a Chemical engineering. $3 560457
650 4 $a Cellular biology. $3 3172791
690 $a 0542
690 $a 0379
710 2 $a Northwestern University. $3 1018161
773 0 $t Dissertation Abstracts International $g 55-01B.
790 $a 0163
791 $a Ph.D.
792 $a 1993
793 $a English
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9415697