Enrichment of a single clone from a high diversity library of phage-displayed antibodies by panning withAnopheles gambiae(Diptera: Culicidae) midgut homogenate

2003 ◽  
Vol 93 (1) ◽  
pp. 31-37 ◽  
Author(s):  
G.F. Killeen ◽  
B.D. Foy ◽  
R.H. Frohn ◽  
D. Impoinvil ◽  
A. Williams ◽  
...  

AbstractA high diversity library of recombinant human antibodies was selected on complex antigen mixtures from midguts of femaleAnopheles gambiaeGiles. The library of phage-displayed single chain variable region fragment constructs, derived from β-lymphocyte mRNA of naïve human donors, was repeatedly selected and reamplified on the insoluble fraction of midgut homogenates. Five rounds of panning yielded only one midgut-specific clone, which predominated the resulting antibody panel. InA. gambiae, the epitope was found throughout the tissues of females but was absent from the midgut of males. The cognate antigen proved to be detergent soluble but very sensitive to denaturation and could not be isolated or identified by Western blot of native electrophoresis gels or by immunoprecipitation. Nevertheless, immunohistology revealed that this sex-specific epitope is associated with the lumenal side of the midgut. Severe bottlenecking may limit the utility of phage display selection from naïve libraries for generating diverse panels of antibodies against complex mixtures of antigens from insect tissues. These results suggest that the selection of sufficiently diverse antibody panels, from which mosquitocidal or malaria transmission-blocking antibodies can be isolated, may require improved selection methods or specifically enriched pre-immunized libraries.

1994 ◽  
Vol 269 (13) ◽  
pp. 9533-9538
Author(s):  
S.J. Deng ◽  
C.R. MacKenzie ◽  
J. Sadowska ◽  
J. Michniewicz ◽  
N.M. Young ◽  
...  

Blood ◽  
2002 ◽  
Vol 99 (7) ◽  
pp. 2562-2568 ◽  
Author(s):  
Delin Zhu ◽  
Helen McCarthy ◽  
Christian H. Ottensmeier ◽  
Peter Johnson ◽  
Terry J. Hamblin ◽  
...  

Most patients with follicular lymphoma (FL) have somatically mutated V genes with intraclonal variation, consistent with location in the germinal center site. Using our own and published sequences, we have investigated the frequency of potential N-glycosylation sites introduced into functional VH genes as a consequence of somatic mutation. FL cells were compared with normal memory B cells or plasma cells matched for similar levels of mutation. Strikingly, novel sites were detected in 55 of 70 (79%) patients with FL, compared to 7 of 75 (9%) in the normal B-cell population (P < .001). Diffuse large B-cell lymphoma (DLCL) showed an intermediate frequency (13 of 32 [41%] patients). Myeloma and the mutated subset of chronic lymphocytic leukemia showed frequencies similar to those of normal cells in 5 of 64 (8%) patients and 5 of 40 (13%) patients, respectively. In 3 of 3 random patients with FL, immunoglobulin was expressed as recombinant single-chain Fv inPichia pastoris, and glycosylation was demonstrated. These findings indicate that N-glycosylation of the variable region may be common in FL and in a subset of DLCL. Most novel sites are located in the complementarity-determining regions. VH sequences of nonfunctional VH genes contained few sites, arguing for positive selection in FL. One possibility is that the added carbohydrate in the variable region contributes to interaction with elements in the germinal center environment. This common feature of FL may be critical for tumor behavior.


2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Ali Jaber Naeemah ◽  
Kuan Yew Wong

PurposeThe purpose of this paper is (1) to review, analyze and assess the existing literature on lean tools selection studies published from 2005 to 2021; (2) to identify the limitations faced by previous studies; and (3) to suggest future works that are necessary to facilitate the selection of lean tools.Design/methodology/approachA systematic approach was used in order to identify, collect and select the articles. Several keywords related to the selection of lean tools were used to collect articles from different Scopus indexed journals. Next, the study systematically reviewed and analyzed the selected papers to identify the lean tools' selection method and discussed its features and limitations.FindingsAn analysis of the results showed that previous studies have adopted two types of methods for selecting lean tools. First, there are various traditional methods being used. Second, multi-criteria decision-making (MCDM) methods were commonly used in previous studies, such as the multi-objective decision-making method (MODM), single multi-attribute decision-making (MADM) methods and hybrid (MCDM). Moreover, the study revealed that the lean tools' selection methods in previous studies were based on evaluating the relationship between either lean tools and performance metrics or lean tools and waste, or both.Research limitations/implicationsIn terms of its theoretical value, the study is considered as an extension of the previous researches performed on this topic by determining and analyzing the features of the most selection methods of lean tools. Unlike previous review papers, this review had considered discussing and analyzing the characteristics and limitations of these methods. Section 2.2 of this paper reviewed some of the categories of MCDM methods as well as some of the traditional methods used in the selected previous studies. Section 2.1 of this paper explained the concept of lean management and its application benefits. Further, only three sectors were covered by the previous studies in this review paper. This study also provided recommendations for future research. Therefore, it provided researchers with a good conception of how to conduct the studies on lean tools selection. Besides, knowing the methods used in previous studies can help researchers develop new methods to select the best set of lean tools. That is, this study provided and advanced the existing knowledge base for researchers concerning lean tools selection, especially there is limited availability of review papers on this topic. Moreover, the study showed researchers the importance of the relationship between lean tools and indicators or/and performance indicators to determine the appropriate set of lean tools so that the results of future studies will be more realistic and acceptable.Practical implicationsPractically, manufacturers face a significant challenge when selecting proper lean tools. This study may enhance managers, manufacturers and company's knowledge to identify most of the methods used to choose the best set of lean tools and what are the advantages, disadvantages and limitations of these methods as well as the latest studies that have been adopted in this topic. That means this study can direct companies to prioritize the application of lean tools depending on either the manufacturing performance metrics or/and manufacturing wastes so that they avoid incorrect application of lean tools, which will add more non-value added activities to operations. Therefore companies can decrease the time and cost losses and enhancing the quality and efficiency of the performance. Correctly implementing the best set of lean tools in companies will lead in general to correctly applying lean management in corporations. Therefore, these lean tools can boost the economic aspect of companies and society through reducing waste, improving performance indicators, preserving time and cost, achieving quality, efficiency, competitiveness, boosting employee income and improving the gross domestic product. The correct lean tool selection reduces customer complaints and employee stress and improves work conditions, health, safety and labor wellbeing. Besides, the correct lean tools selection improves materials usage, energy usage, water usage and decreases liquid wastes, solid wastes and air emissions. As a result, the right selection of lean tools will have positive effects on both the environment and society. The study may also encourage manufacturers and researchers to adopt studies on lean tools selection in small- and medium-sized companies because the study referred to the importance and participation of these kinds of companies in a large proportion of the economy of developing countries. Further, the study may encourage some countries that have not previously adopted this type of study, academically and industrially to conduct lean tools selection studies.Social implicationsAs mentioned previously, the correct lean tool selection reduces customer complaints and employee stress and improves work conditions, health, safety and labor wellbeing. The proper lean tools selection improves materials usage, energy usage, water usage and decreases liquid wastes, solid wastes and air emissions. As a result, the right choice of lean tools will positively affect both the environment and society.Originality/valueThe study expanded the efforts of previous studies concerning lean management features. It provided an accurate review of most lean tools selection studies published from 2005 to 2021 and was not limited to the manufacturing sector. It further identified and briefly described the selection methods concerning lean tools adopted in each paper.


2018 ◽  
Vol 02 (03) ◽  
pp. 169-183
Author(s):  
Sharath Kumar G G ◽  
Chinmay Nagesh

AbstractAppropriate patient selection and expedient recanalization are the mainstay of modern management of acute ischemic stroke (AIS). Only a minority of patients (7–15%) of patients are eligible for endovascular therapy. Patient selection may be time based or perfusion based. Central to both paradigms is the selection of a patient with a small core, a significant penumbra that can be differentiated from areas of oligemia. A brief review of patient selection methods is presented. Endovascular thrombectomy techniques using stentrievers or aspiration catheters have now become the treatment of choice for AIS with large vessel occlusion. A range of devices, each with its own advantages and disadvantages, are available in the market for the neurointerventionist to choose. Techniques vary between devices and between operators, but standardization and protocolization are important within each center. Complications must be anticipated to be avoided. Once reperfusion is achieved, outcomes must be safeguarded with competent postprocedure management to prevent secondary brain injury. These aspects are reviewed in this article.


2019 ◽  
Vol 21 (3) ◽  
pp. 332-339 ◽  
Author(s):  
Jong-Won Chung ◽  
Beom Joon Kim ◽  
Han-Gil Jeong ◽  
Woo-Keun Seo ◽  
Gyeong-Moon Kim ◽  
...  

Biomolecules ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 517
Author(s):  
Seohee Chang ◽  
Soohyun Kim ◽  
Jerome Han ◽  
Suji Ha ◽  
Hyunho Lee ◽  
...  

Phage display is one of the most frequently used platform technologies utilized to screen and select therapeutic antibodies, and has contributed to the development of more than 10 therapeutic antibodies used in the clinic. Despite advantages like efficiency and low cost, it has intrinsic technical limitations, such as the asymmetrical amplification of the library after each round of biopanning, which is regarded as a reason for it yielding a very limited number of antigen binders. In this study, we developed a high-throughput single-clonal screening system comprised of fluorescence immunoassays and a laser-driven clonal DNA retrieval system using microchip technology. Using this system, from a single-chain variable fragment (scFv) library displayed on phages with a complexity of 5.21 × 105 harboring random mutations at five amino acid residues, more than 70,000 clones—corresponding to ~14% of the library complexity—were screened, resulting in 78 antigen-reactive scFv sequences with mutations restricted to the randomized residues. Our results demonstrate that this system can significantly reduce the number of biopanning rounds, or even eliminate the need for this process for libraries with lower complexity, providing an opportunity to obtain more diverse clones from the library.


2010 ◽  
Vol 58 (9) ◽  
pp. 5387-5391 ◽  
Author(s):  
Shuang Wen ◽  
Xiao Zhang ◽  
Yuan Liu ◽  
Qingqing Zhang ◽  
Xianjin Liu ◽  
...  

1998 ◽  
Vol 64 (7) ◽  
pp. 2490-2496 ◽  
Author(s):  
Kai Koo ◽  
Peggy M. Foegeding ◽  
Harold E. Swaisgood

ABSTRACT The variable-region genes of monoclonal antibody againstBacillus cereus spores were cloned from mouse hybridoma cells by reverse transcription-PCR. The heavy- and light-chain variable-region genes were connected by a 45-base linker DNA to allow folding of the fusion protein into a functional tertiary structure. For detection of protein expression, a 10-amino-acid strep tag (biotin-like peptide) was attached to the C terminus of recombinant antibody as the reporter peptide. The single-chain antibody construct was inserted into the expression vector and expressed in Escherichia coliunder the control of the T7 RNA polymerase-T7 promoter expression system. The expressed single-chain antibody was detected on Western blots by using a streptavidin-conjugated enzyme system. This small recombinant antibody fragment (ca. 28,000 Da by calculation) hadB. cereus spore binding ability and antigen specificity similar to those of its parent native monoclonal antibody.


Blood ◽  
1994 ◽  
Vol 83 (11) ◽  
pp. 3279-3288 ◽  
Author(s):  
RE Hawkins ◽  
D Zhu ◽  
M Ovecka ◽  
G Winter ◽  
TJ Hamblin ◽  
...  

Abstract Idiotypic determinants on neoplastic B cells could provide tumor antigens for vaccination of patients with B-cell tumors. Because this approach requires an individual vaccine for each patient, simple methods for obtaining idiotypic antigen are desirable. Using polymerase chain reaction (PCR) with family-based V-gene and J-region primers, the variable region genes of heavy and light chains (VH and VL) of Ig have been obtained from biopsy material from 13 patients with B-cell tumors. In each case, analysis of random clones derived from the PCR product showed repeated, clonally-related sequences, whereas normal lymphoid tissue generated no repeated sequences. In 3/3 cases, the repeated sequences were found to be the same as those in a tumor-derived hybridoma. Mutational patterns in the V-genes differed among the tumors, with follicular lymphoma tending to be more highly mutated. The individual VH and VL sequences have been assembled with a flexible linker sequence to encode single-chain Fv (scFv). The scFv sequences can be cloned into bacterial expression vectors to produce protein, or into vectors suitable for direct vaccination using naked DNA. In a model system, expressed scFv protein retained all idiotypic determinants defined by a panel of five anti-idiotypic monoclonal antibodies (MoAbs). Similarly, expressed scFv proteins from two patients were shown to react with anti-idiotypic antibodies. This approach allows production of potential vaccines from surgical biopsies within 2 to 3 weeks.


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