Antibody production with site-specific non-natural amino acid incorporation for generation of antibody drug conjugates

Ambrx’s mammalian expression platform (EuCODE) enables site-specific incorporation of non-natural amino acids into antibodies. This EuCODE technology provides a means for stable payload linkages at defined sites and with a defined DAR of 2 for antibody drug conjugate (ADC) generation. While the ability to control the DAR and payload site can provide an advantage to an ADC, the incorporation of the non-natural amino acid into the antibody heavy chain introduces a unique challenge for antibody production. To achieve high production of monoclonal antibodies (mAbs) containing non-natural amino acids, we engineered a CHO-K1 cell line which stably contains Ambrx’s patented tRNA and RNA synthetase system components. Then this engineered CHO-K1 cell line was transfected with the gene of interest to generate stable cell lines producing mAbs containing Ambrx’s non-natural amino acid at selected sites. Methodology for vector design (single vector containing both the heavy chain and light chain versus two separate vectors) was investigated for high producing clone generation. In order to achieve manufacturability of the technology, a fed batch process was developed to achieve high titer. Basal media, feed, and feeding strategy of feed supplements and the non-natural amino acid were studied. Furthermore, we investigated how media components affect the efficiency of non-natural amino acid incorporation in mAbs

The Switch: Understanding the Perception of African American Linguistic Repertoires

The purpose of this qualitative study is to conduct research to gain knowledge of the perception of African American adults who use standard English and African American Vernacular English or feel that they must code-switch between the two languages. “The goal is to investigate the attitudes and ideologies that African American Adults have toward AAVE and what role the two languages play in their personal identity. This research will obtain a better understanding of how our use of language can influence our self-concept and identity. Cultural influences can influence our language and how we conceptualize who we are and where we come from. This research intends to examine dialect codeswitching and its place in the dynamic of interracial communication outwardly from the African American community. The study will be extremely beneficial for professional communicators, in understanding why African American code-switching in the workplace, educational, or business settings. Learning why African Americans code switch can help build stronger relationships throughout the workplace, reinforce Diversity, Equity & Inclusion initiatives, and add clarity across companies to create a high-performance culture when communicating with their managers, heads of departments, employees, clients, or stakeholders. Furthermore, this research aims to discover if the black community considers these efforts successful in cultural preservation or representation of African American English dialect as a class of prestige. Finally, identifying these linguistic and cultural differences through a literature review will help to add reader context to the discussion of linguistic inequality and methods of linguistic oppression

Perceived Strengths of the Georgia 4-H Program: A Delphi Analysis

The Georgia 4-H program is the largest youth organization in the state andempowers youth participants to become autonomous and productive members ofsociety. 4-H involvement has been linked to positive youth development outcomes, including leadership and vocational skills, increased self-confidence, and meaningful relationships. The focus of this study was to identify key strengths ofthe 4-H program, as indicated by Foundation Advisory Board members (some ofwhom were 4-H alumni), to guide future programming and curriculum efforts.Strengths were examined through an appreciative inquiry using organizationalmeasures, including positive youth development as conceptualized by T he Five C’s Model (Lerner et al., 2000) and positive organizational behavior as conceptualized by the C . H.O .S.E. characteristics (Luthans, 2002). Through a modified Delphi approach, an expert panel (n =31) identified and reached consensus regarding perceived strengths of the Georgia 4-H program. Eighty-seven items were retained after the Delphi process and were analyzed using the constant comparative method (CCM). Of these 87 items, 41 received a unanimous individual consensus rating. Seven themes emerged following the CCM analysis, including 4-H Organization – General; 4-H Organization – Leadership andSupport; Leadership and Professional Development; Character and Life SkillsDevelopment; Youth Development; Diversity, Accessibility, and Inclusion; and Programming and Opportunitie

Identifying Respondent Group Representation in Extension Capacity Assessments: A Meta-Synthesis of the Literature and a Primary Study

A key factor in determining the future of agricultural extension efforts is ensuring that the voices of those who need to be heard are represented at all stages of the decision-making process. As agricultural extension becomes increasingly globalized, it is critical that the diversity of voices represented within capacity assessments likewise increases. Using two distinct approaches, the present study attempts to address a current gap within the extension literature specifically related to extension assessment respondent groups. First, 97 extension related assessment manuscripts were identified during a literature review and analyzed for respondent group. The results indicated most studies included only one respondent group. Among these assessments Clientele and Beneficiaries and Formal Power Roles were the respondent group categories most frequently examined. Next, a primary study was conducted to identify which respondent groups should be represented in capacity assessment according to agricultural extension experts. The panelists had the highest level of agreement regarding the inclusion of extension clientele and beneficiaries within capacity assessments. However, panelists agreed that representation from outside influences and formal power roles were also important to include in the capacity assessment process. The results indicate extension networks should purposively include a diverse set of respondents when conducting assessments to ensure a comprehensive perspective is represented

Higher Education Leadership Development During the COVID-19 Pandemic: An Exploration of Online Learner Readiness

The COVID-19 pandemic precipitated profound disruptions across the higher education sector as institutions were forced to restructure entire systems and operate with significantly reduced resources. Most notably, many institutions were forced to transition to fully virtual instruction. The present study examined adult leadership development program participants’ perceptions of online learning readiness during the transition to a fully virtual training environment precipitated by the COVID-19 pandemic. A census of the 2020 LEAD21 class was taken, and perceptions of online learning readiness were collected via a retrospective pre- and post-test. Descriptive and inferential statistics were used. Respondents had the highest levels of agreement with computer and Internet self-efficacy and the lowest levels of agreement with learner control in an online context. A paired t-test was conducted to analyze the difference in perceptions of online learning readiness post-training and retrospective pre-training. A statistically significant increase was observed for overall online learning readiness, as well as for computer and Internet self-efficacy, learner control, motivation for learning, Internet communication self-efficacy, and self-directed learning. An implication from this finding is that the transition to fully virtual training resulted in increased online learning readiness across all dimensions. With new strains of COVID-19 emerging and the potential for ongoing restrictions for social interaction, online learning will continue to be an important aspect of the educational process. It is vital that higher education leaders consider individuals’ readiness to effectively engage in online training and instruction. Implications and recommendations for future research in practice in international contexts are provided

Information and Communication Technology Use Capacity Within Extension Networks: Development and Preliminary Validation of an Empirical Scale

Advancing information and communication technologies (ICTs) has become central to international agricultural and extension development efforts. ICTs are crucial in facilitating information transfer, ensuring stakeholder access to information, and increasing the decision-making capacity of smallholder farmers. The research presented here introduces an instrument developed to quantify perceptions of ICT use capacity within international extension networks. The aggregate scale was verified for content validity, response process validity, internal structure validity, and consequential validity informing its use. The instrument was administered to network members (n = 122) associated with the Global Forum for Rural Advisory Services. An exploratory factor analysis (EFA) was conducted with measures of correlation and reliability analyzed. Six factors were extracted and analyzed further. The resulting Perceptions of ICT Use scale and factors can be used as reliable instruments for quantifying perceptions of ICT use capacity, enhancing international extension network needs assessments, and informing policies and practices which maximize ICT capacity

Identifying Respondent Group Representation in Extension Capacity Assessments: A Meta-Synthesis of the Literature and a Primary Study

A key factor in determining the future of agricultural extension efforts is ensuring that the voices of those who need to be heard are represented at all stages of the decision-making process. As agricultural extension becomes increasingly globalized, it is critical that the diversity of voices represented within capacity assessments likewise increases. Using two distinct approaches, the present study attempts to address a current gap within the extension literature specifically related to extension assessment respondent groups. First, 97 extension related assessment manuscripts were identified during a literature review and analyzed for respondent group. The results indicated most studies included only one respondent group. Among these assessments Clientele and Beneficiaries and Formal Power Roles were the respondent group categories most frequently examined. Next, a primary study was conducted to identify which respondent groups should be represented in capacity assessment according to agricultural extension experts. The panelists had the highest level of agreement regarding the inclusion of extension clientele and beneficiaries within capacity assessments. However, panelists agreed that representation from outside influences and formal power roles were also important to include in the capacity assessment process. The results indicate extension networks should purposively include a diverse set of respondents when conducting assessments to ensure a comprehensive perspective is represented

CRISPR-Cas9 mediated cell line engineering of apoptosis pathways increases antibody expression with site-specific modifications for antibody drug conjugation

New generation of antibody drug conjugates (ADCs) have expanded the repertoire of antibody drugs in the clinic and the market for cancer and inflammation indications by using highly stable linkers to attach potent small-molecule drug to various targeting antibodies. The drug and site of drug linkage to the antibody can have profound impact on the physiochemical properties and pharmacological profile of the ADC. Ambrx has developed a technology, Eukaryotic Chemical Orthogonal Directed Engineering (EuCODE), which allows non-natural amino acids with diverse physicochemical and biological properties to be genetically encoded and site-specifically incorporated into proteins/antibodies in mammalian cells. The non-natural amino acid provides a handle for the attachment of a small-molecule drug to generate homogenous ADC with a defined Drug-to-Antibody Ratio (DAR). To establish a CHO expression system for high production of monoclonal antibodies (mAbs) containing non-natural amino acids, we successfully generated a EuCODE platform cell line stably expressing engineered amber suppressor tRNA and its cognate tRNA synthetase specific for non-natural amino acid para-acetyl phenylalanine (pAF). When transfected with antibody of interest engineered with amber nonsense codon (TAG) at selected sites suitable for drug conjugation, this EuCODE platform cell line generates stable cell lines producing pAF containing mAbs for site-specifically conjugated ADC. In order to improve production titers of pAF containing antibody and achieve a robust platform, the platform cell line and stable cell lines were further evolved using CRISPR/Cas9 genome editing technology to sequentially knock out selected genes in glutamine synthesis and apoptosis pathways to improve selection efficiency and prevent loss of viable cell mass in production cultures, respectively. Inhibition of apoptosis pathway leads to dramatic increase in viable cell mass and results in extended production time and increased productivity. Phenotypic and genetic properties of these CRISPR engineered cell lines and product quality of the antibody will be discussed in the context of using the platform to develop a commercial manufacturing cell line

Development and Validation of an Empirical Instrument to Measure Organizational and Institutional Functioning Capacity Within International Extension Networks

International extension networks provide vital services to stakeholders using a variety of organizational structures. To continue offering services desired by stakeholders, it is important that both public and private extension networks develop effective organizational functioning capacities. Despite this need, an instrument to empirically examine the perceived capacities of effective international extension network organizational and institutional functioning was unavailable in the literature. The present study developed and validated the International Extension Network Organizational Functioning (IENOF) scale. Scale development included validation of content, response process, internal structure, and consequential utility. Structural analysis indicated the 21-items loaded on four factors, including: network structure and relevance, network management and activities, network strategy and guidance, and network leadership. The results of the study indicate the IENOF may provide an empirical instrument to examine the capacities of international extension network organizational functioning. Furthermore, the analysis provides insights into the nuance and unique characteristics associated with international extension network organizations. Replication studies and further statistical analyses are recommended to examine the nature of the instrument and associated outcome variables of interest. From a practical perspective the IENOF is recommended to be considered as an input to the theory of co-innovation to aid international extension networks in fostering a culture of innovation. Utilizing IENOF insights to inform the theory of co-innovation should better enable extension networks to provide services and programming that meet the needs of network stakeholders

Development and Validation of the International Extension Network Advocacy Capacity Scale

Extension is sometimes viewed globally as less important than academic research and innovation, even though these networks are crucial to agricultural development. Using Lewin’s (1947) organizational change theory as a conceptual background, an instrument to measure advocacy capacity within international extension networks was developed. The resulting scale was analyzed for content validity, response process validity, internal structure validity, and consequential validity. An exploratory factor analysis (EFA) was performed on the proposed scale. The underlying structure of the scale was found to load onto five factors. The five extracted factors were labelled: 1) network support of advocacy, 2) network integration of advocacy, 3) network influence in advocacy, 4) network visibility in advocacy, and 5) network advocacy performance. The overall International Extension Network Advocacy Capacity (IENAC) scale, and associated subscales were all found to have acceptable levels of validity