scholarly journals A cell suspension based uptake method to study high affinity glucosinolate transporters

Plant Methods ◽  
2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Deepti M. Nambiar ◽  
Juhi Kumari ◽  
Gulab C. Arya ◽  
Amarjeet K. Singh ◽  
Naveen C. Bisht
Keyword(s):  
Cell Suspension ◽  
High Affinity ◽  
A Cell ◽  
Uptake Method

Combination of ribosome and phage display for fast selection of high affinity VHH antibody fragments

2019 ◽  
pp. 27-33
Author(s):  
YuE Kravchenko ◽  
SV Ivanov ◽  
DS Kravchenko ◽  
EI Frolova ◽  
SP Chumakov
Keyword(s):  
Phage Display ◽  
Selection Procedure ◽  
Free System ◽  
Phage Displays ◽  
High Affinity ◽  
Binding Domains ◽  
A Cell ◽  
Vhh Antibodies ◽  
Efficiency Of Selection ◽  
Selection Of

Selection of antibodies using phage display involves the preliminary cloning of the repertoire of sequences encoding antigen-binding domains into phagemid, which is considered the bottleneck of the method, limiting the resulting diversity of libraries and leading to the loss of poorly represented variants before the start of the selection procedure. Selection in cell-free conditions using a ribosomal display is devoid from this drawback, however is highly sensitive to PCR artifacts and the RNase contamination. The aim of the study was to test the efficiency of a combination of both methods, including pre-selection in a cell-free system to enrich the source library, followed by cloning and final selection using phage display. This approach may eliminate the shortcomings of each method and increase the efficiency of selection. For selection, alpaca VHH antibody sequences suitable for building an immune library were used due to the lack of VL domains. Analysis of immune libraries from the genes of the VH3, VHH3 and VH4 families showed that the VHH antibodies share in the VH3 and VH4 gene groups is insignificant, and selection from the combined library is less effective than from the VHH3 family of sequences. We found that the combination of ribosomal and phage displays leads to a higher enrichment of high-affinity fragments and avoids the loss of the original diversity during cloning. The combined method allowed us to obtain a greater number of different high-affinity sequences, and all the tested VHH fragments were able to specifically recognize the target, including the total protein extracts of cell cultures.

Experimental study of diffusion-based extraction from a cell suspension

2008 ◽  
Vol 5 (4) ◽  
pp. 529-540 ◽  
Author(s):  
Clara Mata ◽  
Ellen K. Longmire ◽  
David H. McKenna ◽  
Katie K. Glass ◽  
Allison Hubel
Keyword(s):  
Experimental Study ◽  
Cell Suspension ◽  
A Cell

scholarly journals The use of a slide spinner in the analysis of cell dispersion.

1976 ◽  
Vol 24 (1) ◽  
pp. 11-15 ◽  
Author(s):  
R C Wolley ◽  
H M Dembitzer ◽  
F Herz ◽  
K Schreiber ◽  
L G Koss
Keyword(s):  
Cell Suspension ◽  
Single Cells ◽  
Cell Loss ◽  
Optimum Conditions ◽  
Isolated Cells ◽  
Cell Dispersion ◽  
Cervical Cancer Cells ◽  
A Cell ◽  
Human Cervical Cancer Cells ◽  
Human Cervical Cancer

A simple and reliable method of determining the degree of dispersion of a cell suspension has been developed using the Perkin-Elmer Uni-Smear Spinner. Optimum conditions regarding rate and duration of spin, etc., were first ascertained using dispersed cell cultures including human cervical cancer cells as well as gynecologic samples. After spinning, single cells in suspension appeared as isolated cells on the slides. Cell aggregates, on the other hand, remained together. Therefore, the distribution of cells in various sized aggregates could be easily quantitated and the slides retained for future review. This method was used to evaluate the dispersing effects of trypsin, ethylenediaminetetraacetate and and syringing human on human gynecology samples obtained by routine cervical scrapes. None of the dispersion methods has, so far, produced an adequate monodispersed cell suspension without unacceptable cell loss.

Accumulation and Biotransformation of Chromenes and Benzofurans in a Cell Suspension Culture ofAgeratina adenophora

Planta Medica ◽  
1987 ◽  
Vol 53 (05) ◽  
pp. 488-492 ◽  
Author(s):  
Peter Proksch ◽  
Ludger Witte ◽  
Victor Wray ◽  
Ines Rahaus
Keyword(s):  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
A Cell

Analyse de la morphogenèse du pied des Oiseaux à l'aide de mélanges cellulaires interspécifiques

Development ◽  
1973 ◽  
Vol 29 (1) ◽  
pp. 175-196
Author(s):  
Par Marie-Paule Pautou
Keyword(s):  
Cell Suspension ◽  
The Other ◽  
Composite Type ◽  
A Cell ◽  
Species Specific ◽  
Host Embryo

Morphogenesis of the feet of birds, studied in limbs developed from reaggregated heterospecific mesoderm Experiments were undertaken to determine whether species-specific characters of chick and duck mesodermal leg-bud cells are retained after dissociation and reaggregation in homoand heterospecific mixtures. Prospective zeugopod and autopod mesoderm from chick and/or duck leg buds were isolated, dissociated into a cell suspension and pelleted by centrifugation. The reaggregated mesoderm was packed into a leg-bud ectodermal jacket; the recombined leg bud was then grafted on the wing stump of a host embryo. Recombinants whose mesoderm was a homospecific reaggregate developed into typical chick or duck leg parts according to the specific origin of the mesodermal component; the feet of nearly all these legs lacked antero-posterior polarity. Recombinants containing heterospecific reaggregates were also capable of forming reasonably organized leg structures. The foot was not, as a rule, of the specific type expected of the majority component. In a mixture of 75% chick mesoderm cells and 25% duck mesoderm cells, the feet which developed were either of chick type or of composite chick/duck type, where typical chick areas were next to typical or aberrant (steganoid) duck areas. When the ratio was reversed (25% chick, 75% duck), the majority of the feet were again of chick type or of composite chick/duck type, the typical duck phenotype being exceptional. Even in a mixture of 10% chick cells and 90% duck cells, duck-type feet were not obtained. They were all of composite type: half of their interdigital zones were of chick type, the other half were occupied, in most cases, by underdeveloped, indented webbing or by one or several discrete flaps, and, in a few cases, by normal webbing. The vast majority of the feet developed from heterospecific mesoderm were characterized by the profusion of the toes, which were not polarized along the a–p axis.

scholarly journals Lupeol acetate production and antioxidant activity of a cell suspension culture from Cnidoscolus chayamansa leaves

2019 ◽  
Vol 125 ◽  
pp. 30-38 ◽  
Author(s):  
M.Z. Pérez-González ◽  
A. Nieto-Trujillo ◽  
G.A. Gutiérrez-Rebolledo ◽  
I. García-Martínez ◽  
M.E. Estrada-Zúñiga ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
Acetate Production ◽  
A Cell ◽  
Lupeol Acetate ◽  
Cnidoscolus Chayamansa

Preparation Of Factor IX-Deficient Human Plasma Using RFF-IX/1 Monoclonal Antibody

1981 ◽  
Author(s):  
F Rotblat ◽  
A H Goodall ◽  
G Janossy ◽  
G Kemble ◽  
D P O’Brien ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cell Line ◽  
Factor Ix ◽  
Hybrid Cells ◽  
Spleen Cells ◽  
High Affinity ◽  
A Cell ◽  
A Site ◽  
Monoclonal Cell Line ◽  
Mouse Myeloma

A cell line that secretes a monoclonal antibody to factor IX has been produced by fusing spleen cells from a mouse that had been hyper immunised to purified factor IX with mouse myeloma cells (line P3-NSI/I-Ag4-1). Hybrid cells were selected and a monoclonal cell line has been established in culture. This cell line secretes an IgGl(k) antibody (RFF-IX/1) with high affinity for a site related to the coagulant function of factor IX.Monoclonal antibody was partially purified from ascitic fluid from mice implanted with the RFF-IX/1 secreting cells by precipitation at 50% saturation with ammonium sulphate. This fraction has typically 630 NIH units/ml anti IX activity and 13.5 mg/ml protein. It was coupled to cyanogen bromide activated Sepharose 2B in the ratio of 9 mg. protein/1 ml gel. A column containing 10 ml of this gel removed all the assayable factor IX from the first 280 ml of normal ci.trated plasma that was passed over it. After that volume small amounts of factor IX could be detected in the effluent. Subsequently 10-20% of the factor IX activity adsorbed could be recovered by eluting the column with 3 M potassium iodide.Immuno-affinity depleted plasma could be used as substrate in a one-stage factor IX assay under routine laboratory conditions and was undistinguishable for that purpose from severe Christmas disease plasma.

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