IT Strategy for Construction Companies: A Pragmatist’s Vision

Abstract: Leading constructors will alter the economic model in order to take advantage of IT opportunities in the coming years. Strategic business leaders aren’t interested in playing with computer tools that nibble at the margins of individual productivity; they seek the substantial savings in costs and improvements in service that have been won in manufacturing, retail, and financial services industries. These victories are achieved by using the power of technology to streamline and improve entire supply chains; not just to automate processes within the walls of one firm. The construction industry is currently organized on an “every man for himself ” business basis: each firm holds its information within its walls. Therefore, the compelling IT strategies in construction and engineering will incorporate economic incentives that overcome this barrier and make it worthwhile for individual actors to aid the whole supply chain. What are these tools? How might the incentives be structured? The marriage of technology tools and business incentives is the core of a strategic vision for information technology in construction for the next decade. The author is Chairman of the George B. H. Macomber Company, a large regional general contractor based in Boston. He was founder and CEO of Collaborative Structures, Inc, a pioneering Internet services firm enabling communication on construction projects. He is

Comparison between Prototype Hybrid Capture 3 and Hybrid Capture 2 Human Papillomavirus DNA Assays for Detection of High-Grade Cervical Intraepithelial Neoplasia and Cancer

We compared the performance of a prototype version of the Hybrid Capture 3 (HC3) human papillomavirus (HPV) DNA assay to the current generation Hybrid Capture 2 (HC2) assay, both of which target 13 oncogenic HPV types, for the detection of cervical intraepithelial neoplasia grade 3 and cancer (CIN3+) with cervicovaginal lavage specimens collected at enrollment into a 10-year cohort study at Kaiser Permanente (Portland, Oreg.). HC3 results for a risk-stratified sample (n = 4,364) were compared to HC2 results for the entire cohort (n = 20,810) with receiver operating characteristics curves, and the optimal cut points for both tests (relative light units [RLU]/positive control [PC]) for the detection of CIN3+ were determined. Specimens were also tested for HPV16 and HPV18 with separate HC3 type-specific probes. The optimal cut point for detecting CIN3+ was 1.0 RLU/PC for HC2, as previously shown, and was 0.6 RLU/PC for HC3. At the optimal cut points, HC3 and HC2 had similar screening performance characteristics for CIN3+ diagnosed at the enrollment visit. In analyses that included cases CIN3+ at enrollment and those diagnosed during early follow-up, HC3 had nonsignificantly higher sensitivity and equal specificity for the detection of CIN3+ compared to HC2; this increase in sensitivity was primarily the result of increased detection of CIN3+ in women who were 30 years of age or older and were cytologically negative (P = 0.006). We also compared the performance of the hybrid capture tests to MY09/11 L1 consensus primer PCR results (n = 1,247). HC3 was less likely than HC2 to test positive for specimens that tested positive by PCR for any untargeted types (P < 0.001). HC3 was less likely than HC2 to test positive for untargeted PCR-detected single infections with HPV53 (P = 0.001) and HPV66 (P = 0.01). There was good agreement between test positivity by PCR and by single type-specific HC3 probes for HPV16 (kappa = 0.76; 95% confidence interval [CI] = 0.71 to 0.82) and for HPV18 (kappa = 0.73; 95% CI = 0.68 to 0.79). In conclusion, we suggest that HC3 (≥0.6 RLU/PC) may be slightly more sensitive than and equally specific test as HC2 (≥1.0 RLU/PC) for the detection of CIN3+ over the duration of typical screening intervals

High Affinity Humanized Antibodies without Making Hybridomas; Immunization Paired with Mammalian Cell Display and <em>In Vitro</em> Somatic Hypermutation

<div><p>A method has been developed for the rapid generation of high-affinity humanized antibodies from immunized animals without the need to make conventional hybridomas. Rearranged IgH D(J) regions were amplified from the spleen and lymph tissue of mice immunized with the human complement protein C5, fused with a limited repertoire of human germline heavy chain V-genes to form intact humanized heavy chains, and paired with a human light chain library. Completed heavy and light chains were assembled for mammalian cell surface display and transfected into HEK 293 cells co-expressing activation-induced cytidine deaminase (AID). Numerous clones were isolated by fluorescence-activated cell sorting, and affinity maturation, initiated by AID, resulted in the rapid evolution of high affinity, functional antibodies. This approach enables the efficient sampling of an immune repertoire and the direct selection and maturation of high-affinity, humanized IgGs.</p> </div

Discovery and affinity maturation of C5-C345C-specific antibodies.

<p>(<i>A</i>) FACS scattergrams depicting isolation of a C5-C345C-specific population from the sub-library pool containing IGHV3-30-3 and IGHV4-34 (panels I–III), and subsequent affinity maturation of APE777 in strategy B (panels IV–VI). Approximately 5×10⁷ cells were sorted per round. IgG expression is shown on the y-axis and C5-C345C binding on the x-axis. (I) The first sort round (5 µM C5-C345C-Myc) showed little detectable binding; (II) round 4 (100 nM C5-C345C-Myc); and (III) round 9 (3 nM C5-C345C-Myc) which included a room-temperature incubation to increase stringency. Affinity maturation FACS scattergrams of isolated C5-C345C-specific antibody, APE777, are shown for strategy B starting with (IV) round 1 (20 nM C5-C345C-Myc), (V) round 5 (1.5 nM C5-C345C-Dyl-650), and (VI) round 8 (100 pM C5-C345C-Dyl-650). Each sort round consisted of AID-induced SHM and selection of highest antigen binding and antibody expressing cells by FACS. Note emergence of new cell populations that exhibited increased antigen binding relative to the original population over time, and which were able to recognize sequentially decreasing antigen concentrations. (<i>B</i>) Plot comparing stability late (value corresponds to amount of antibody-bound antigen) versus binding late (value corresponds to off-rate, k_d) capture-adjusted report points for Biacore 4000 screen of single cell clone supernatants. Higher values correspond to more antigen bound per unit antibody (binding late) and slower off-rates (stability late). Highlighted points indicate highest affinity C5-C345C binding clones further characterized by Biacore T200 and sequencing. (<i>C</i>) Full kinetic analysis of APE777 binding to C5-C345C (first panel) with a K_D of 200 nM (k_a = 1.4×10⁵ M⁻¹ s⁻¹, k_d = 2.4×10⁻² s⁻¹). Binding to C5 and C5b6 complement proteins was detectible (second and third panels), while APE777 shows no significant binding to the C7 negative control.</p

Summary of observed enriching mutations.

<p>Table of HC and LC point mutation, insertion, and deletion events observed during affinity maturation of APE777. The first column indicates the amino acid mutation, listed with Kabat numbering. The second column indicates whether the mutation is in the HC or LC. Columns three and four show the starting and ending nucleotide sequence surrounding the site of mutation, with the mutated nucleotides underlined. Columns five and six indicate the starting and ending nucleotide for point mutations. The last two columns indicate which strategy the mutation was observed in, and at which round it was enriched. Ten of the 11 point mutations were initiated at G or C nucleotides; one of the 11 was initiated at a T. Seven of the point mutations, the deletion, and one of the insertions occurred at known AID hotspots (WRCH, highlighted in <b>bold</b>) located within CDRs 1, 2, or 3. Seven of the 11 point mutations were nucleotide transitions, and four were nucleotide transversions.</p>1<p>Enrichment observed by Sanger sequencing of 40 HC and LC variable regions post-sort round.</p

Isolation of high-affinity monoclonal antibodies from immunized mice.

<p>Spleen and draining lymph nodes were harvested from mice immunized with the C345C fragment of complement protein C5. After RNA isolation and cDNA synthesis, (D)J diversity was amplified from HCs and cloned into nine human germline variable regions (IGHV 1-2, 1-69, 3-7, 3-23, 3-30, 4-34, 4-59, 5-51, and 6-1). These were paired with a fully human LC library, composed of five germline IGκV regions (IGKV 1-33, 1D-39, 2D-30, 3-20, and 4-1), combined with CDR3 diversity obtained from a pool of normal human donors. Libraries were transfected into HEK 293 c18 cells as four sub-library pools for cell surface display and FACS selection of antigen specific clones. Subsequent affinity maturation by AID-induced mutagenesis and FACS resulted in a high-affinity antibody with demonstrated functional activity.</p