geoscience.net logo
+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Assessing genetic heterogeneity in production cell lines: detection by peptide mapping of a low level Tyr to Gln sequence variant in a recombinant antibody


, : Assessing genetic heterogeneity in production cell lines: detection by peptide mapping of a low level Tyr to Gln sequence variant in a recombinant antibody. Bio/Technology 11(11): 1293-1297

A recombinant antibody directed against the human epidermal growth factor receptor-2 extracellular domain was subjected to detailed structural characterization. Heterogeneity in the heavy chain was demonstrated by recovery of two forms of a tryptic peptide, with either glutamine or the expected tyrosine at residue 376. Subsequent experiments indicated that the Y376Q variant developed during transfection of the antibody heavy and light chain genes into Chinese hamster ovary cells. Levels of the Y376Q variant (range: 27% to 1%) in the purified antibody were inversely proportional to cell age. The established cell line was subcloned and found to be heterogeneous by polymerase chain reaction analysis of cell extracts and protein analysis of the purified antibody. Ten percent of subclones produced high levels of the Y376Q variant while 90% of the subclones produced antibody with only the expected heavy chain sequence. This report demonstrates the utility of peptide mapping as a sensitive tool for assessing genetic heterogeneity of recombinant cell lines.

US$29.90

PMID: 7764191


Other references

Colin, L.; Vandenhoudt, N.; de Walque, Séphane.; Van Driessche, Bît.; Bergamaschi, A.; Martinelli, Vérie.; Cherrier, T.; Vanhulle, C.; Guiguen, A.; David, A.; Burny, Aène.; Herbein, G.; Pancino, G.; Rohr, O.; Van Lint, C., 2011: The AP-1 binding sites located in the pol gene intragenic regulatory region of HIV-1 are important for viral replication. Our laboratory has previously identified an important intragenic region in the human immunodeficiency virus type 1 (HIV-1) genome, whose complete functional unit is composed of the 5103 fragment, the DNaseI-hypersensitive site HS7 and the 5105 fra...

Yang V.C.; Teng C L.C., 1990: An immobilized protamine system for removing heparin in extracorporeal blood circulation. Gebelein, C G (Ed ) Biomimetic Polymers Viii+297p Plenum Press: New York, New York, Usa; London, England, Uk Illus 175-190

Altomare, D.F.; Rinaldi, M.; Troilo, V.L.; Marino, F.; Lobascio, P.; Puglisi, F., 2005: Closed ambulatory lateral internal sphincterotomy for chronic anal fissures. Techniques in Coloproctology 9(3): 248-249

Cacace E.; Mela Q.; Frigerio R.; Sole G.; Olla N.; Carcassi U., 1985: Iron chelating therapy in beta 0 thalassemia intermedia. Aksoy, M And G F B Birdwood (Ed ) Hypertransfusion And Iron Chelation in Thalassaemia; Symposium Held During The Meeting Of The Mediterranean Blood Club, Izmir, Turkey, June 1984 88p Hans Huber Publishers: Berne, Switzerland; Toronto, Ont , Canada Illus Paper 48-61

Nekrasov, V.P.; Rozanov, V.E., 1980: Rare causes of hemorrhages from the small intestine. Data on the main clincal-roentgenological manifestations of acute hemorrhages from benign tumors of the small intestine and Meckel's diverticula in patients are summarized. Despite the absence of specific symptomatology, the source of acute i...

Sawyer, R.T.; Day, B.J.; Fadok, V.A.; Chiarappa-Zucca, M.; Maier, L.A.; Fontenot, A.P.; Silveira, L.; Newman, L.S., 2004: Beryllium-ferritin: lymphocyte proliferation and macrophage apoptosis in chronic beryllium disease. A beryllium (Be)-ferritin adduct containing 270 pm of Be stimulated proliferation of bronchoalveolar lavage (BAL) lymphocytes from subjects with chronic beryllium disease (CBD) at concentrations 5-6 logs lower than the amounts of beryllium sulfate...

Dutton, P.L.; Petty, K.M.; Bonner, H.S.; Morse, S.D., 1975: Cytochrome c2 and reaction center of Rhodospeudomonas spheroides Ga. membranes. Extinction coefficients, content, half-reduction potentials, kinetics and electric field alterations. The reduced minus oxidized extinction coefficients (delta epsilon-red-ox) of reaction center P605 when in the chromatophore is about 20% smaller than in the detergent-isolated state. Presumably the coupling of the reaction center protein to the an...

Mcintosh, S.A.; Simon, R.; Flett, M.M.; Stanton, P.D.; Soukop, M.; Puroshotham, A.D.; Cooke, T.G.; Going, J.J., 1999: Therapeutic implications of cytokeratin IHC evaluation of sentinel lymph node biopsy in breast cancer. Journal of Pathology 189(SUPPL ): 10A

Synge, R.L., 1944: Analysis of a partial hydrolysate of gramicidin by partition chromatography with starch. Biochemical Journal 38(4): 285-294

Monney, N.P-A.; Bally, T.; Giese, B., 2016: Electronic structure of hole-conducting states in polyprolines. Electron transfer over long distances in proteins by a hopping process requires transient relay stations that can harbor charge and spin for a short time span. Certain easily oxidizable or reducible side chains may assume that role, but it has bee...