Formulation Considerations for Autologous T Cell Drug Products.

Genetically modified autologous T cells have become an established immunotherapy in the fight against cancer. The manufacture of chimeric antigen receptor (CAR) and αß-T cell receptor (TCR) transduced T cells poses unique challenges, including the formulation, cryopreservation and fill-finish steps, which are the focus of this review. With an increasing number of marketing approvals for CAR-T cell therapies, comparison of their formulation design and presentation for administration can be made. These differences will be discussed alongside the emergence of automated formulation and fill-finish processes, the formulation design space, Monte Carlo simulation applied to risk analysis, primary container selection, freezing profiles and thaw and the use of dimethyl sulfoxide and alternative solvents/excipients as cryopreservation agents. The review will conclude with a discussion of the pharmaceutical solutions required to meet the simplification of manufacture and flexibility in dosage form for clinical treatment.

Comparison between the different methods developed for determining the onset of the LH surge in urine during the human menstrual cycle.

PURPOSE: To determine whether an optimal method exists for the detection of the luteinising hormone (LH) surge onset in research datasets of urinary hormonal profiles of menstrual cycles. METHODS: The scientific literature was searched to compare published methodologies for detection of the LH surge onset in urine. Their performance was tested using complete hormonal profiles from 254 ovulatory cycles from 227 women attempting pregnancy (normal regular menstrual cycles; no known infertility). RESULTS: Three major methodologies to determine the onset of the LH surge in urine were identified. The key difference between these methods is how the cycle days that contribute to LH baseline assessment are determined: using fixed days (method #1), based on peak LH day (method #2), based on a provisional estimate of the LH surge (method #3). Method #1 requires no prior cycle information, whereas methods #2 and #3 need to consider complete cycle data. The most reliable method for calculation of baseline LH was using 2 days before the estimated surge day, plus the previous 4/5 days. CONCLUSIONS: Different methods for identification of the urinary LH surge can provide very different determinations of LH surge day, thus care must be taken when comparing between studies that apply different methodologies. The optimal method for determining the onset of the LH surge in urine requires retrospective estimation of day of LH surge to identify the most appropriate part of the cycle to consider as the baseline. This method can be adopted for application in population studies.

Development of the first urinary reproductive hormone ranges referenced to independently determined ovulation day.

BACKGROUND: Urinary hormone level analysis provides valuable fertility status information; however, previous studies have not referenced levels to the ovulation day, or have used outdated methods. This study aimed to produce reproductive hormone ranges referenced to ovulation day determined by ultrasound. METHODS: Women aged 18-40 years (no reported infertility) collected daily urine samples for one complete menstrual cycle. Urinary luteinising hormone (LH), estrone-3-glucuronide (E3G, an estradiol metabolite), follicle stimulating hormone (FSH) and pregnanediol-3-glucuronide (P3G, a progesterone metabolite) were measured using previously validated assays. Volunteers underwent trans-vaginal ultrasound every 2 days until the dominant ovarian follicle size reached 16 mm, when daily scans were performed until ovulation was observed. Data were analysed to create hormone ranges referenced to the day of objective ovulation as determined by ultrasound. RESULTS: In 40 volunteers, mean age 28.9 years, urinary LH surge always preceded ovulation with a mean of 0.81 days; thus LH is an excellent assay-independent predictor of ovulation. The timing of peak LH was assay-dependent and could be post-ovulatory; therefore should no longer be used to predict/determine ovulation. Urinary P3G rose from baseline after ovulation in all volunteers, peaking a median of 7.5 days following ovulation. Median urinary peak E3G and FSH levels occurred 0.5 days prior to ovulation. A persistent rise in urinary E3G was observed from approximately 3 days pre- until 5 days post-ovulation. CONCLUSIONS: This study provides reproductive hormone ranges referenced to the actual day of ovulation as determined by ultrasound, to facilitate examination of menstrual cycle endocrinology.

Comparison of analytical sensitivity and women's interpretation of home pregnancy tests.

BACKGROUND: Internationally there are no defined standards for expressing the performance of home-based pregnancy tests, nor any pre-defined accuracy standard requirements. The aim of this study was to evaluate the accuracy of a selection of home-based pregnancy tests relative to their packaging/product insert claims. METHODS: Eight home-based pregnancy tests were evaluated using human chorionic gonadotrophin (hCG) urine standards (0, 15, 25 mlU/mL). Testing was performed by a technician and results were read by a technician and a panel of consumer volunteers (each blinded to the expected result) and compared with the expected result based on the manufacturer's claimed accuracy. Volunteers also completed questionnaires relating to various device attributes. RESULTS: The overall agreement between the technician reading and expected reading from the hCG concentration was >90% for the Clearblue® DIGITAL, Clearblue® PLUS, Confirme® Plus, David® and Haus™ tests, and approximately 80% for Predictor® Early; agreement was <50% for Femitest® Jet Ultra and Cyclotest® Early tests. RESULTS were available from 72 volunteers (aged 18-45 years). Overall the percentage agreement between volunteer result and expected result was >95% for Clearblue DIGITAL and Clearblue PLUS tests; agreement for all other tests was <75% (lowest were Cyclotest Early and Femitest Jet Ultra, 33.0% and 39.4%, respectively). The Clearblue DIGITAL test was scored most highly by volunteers in the questionnaires. CONCLUSIONS: Many home-based pregnancy tests commonly used by women are not as accurate as their packaging information claims. International test standards which define appropriate performance characteristics for home pregnancy tests are urgently required.

Accuracy of a home-based device for giving an early estimate of pregnancy duration compared with reference methods.

OBJECTIVE: To assess a home pregnancy test's accuracy to concurrently detect pregnancy and determine pregnancy duration. DESIGN: Multicenter, prospective study. SETTING: Study sites in the United States. PATIENT(S): Women actively attempting to conceive who have menstrual bleeds (18-45 years). INTERVENTION(S): Volunteers collected early morning urine samples (three or fewer menstrual cycles). Pregnant volunteers underwent ultrasound dating scans. Ovulation day (LH surge +1 day) during pregnancy-resulting cycles was determined by quantitative measurement of LH. Random urine samples were tested with the hCG-measuring pregnancy test from 4 days before the expected period until 4 weeks later. MAIN OUTCOME MEASURE(S): A home pregnancy test's accuracy in determining pregnancy duration compared with ultrasound and ovulation day. RESULT(S): Agreement between pregnancy test results and time since ovulation was 93% (confidence interval [CI], 91.5-94.4). Agreement with ultrasound was dependent on the formula: there was 99% agreement when calculated with adjustment for Hadlock formula bias (Pexsters; CI, 98.2-99.4) or using a nonbias formula (Wu; CI, 98.6-99.6), when ultrasound error was accommodated. Agreement was lower when bias/measurement errors were not accounted for (Wu, 86%, CI, 83.9-88; Hadlock, 80.8, CI, 78.2-83.3). CONCLUSION(S): This home pregnancy test provides an accurate estimation of pregnancy duration in weeks categories, 1-2, 2-3, 3+ weeks since ovulation, thereby showing utility in dating pregnancy.

Human chorionic gonadotropin as a measure of pregnancy duration.

OBJECTIVE: To compare gestational age (GA) estimates in early pregnancy, determined by last menstrual period (LMP), human chorionic gonadotropin (hCG) concentration, ultrasound crown-rump length (Hadlock formula), and ovulation day (luteinizing hormone surge plus 1day). METHODS: Female volunteers seeking to conceive (at 5 US sites) collected daily early-morning urine for up to 3 menstrual cycles. Pregnant women underwent ultrasound dating scans. Conception cycle urine was quantitatively assessed for luteinizing hormone and hCG. Summary statistics for GA using each reference method were determined (n=131). RESULTS: Correlation between GA determined by ultrasound and ovulation day was excellent (maximum difference 10days); however, pregnancies dated by ultrasound were 3days advanced. The difference between LMP estimates and estimates based on ovulation day or ultrasound was 9 and 12days, respectively. A uniform rise in hCG on each day of pregnancy was seen using all reference methods. The accuracy of hCG measurement in determining the week since conception was more than 93%. CONCLUSION: Methods for establishing pregnancy duration vary in their accuracy and their GA estimates. The rise in hCG concentration in early pregnancy is uniform and therefore hCG levels provide the most accurate, early estimation of GA in single, viable pregnancies. ClinicalTrials.gov:NCT01077583.

Comparison of volunteers' experience of using, and accuracy of reading, different types of home pregnancy test formats.

BACKGROUND: Women suspecting pregnancy need an accurate result when they conduct a home pregnancy test. A variety of tests are available from simple professional style strips to midstream tests with a digitally displayed result. However, it is not known whether all these formats can be used and read correctly by untrained women. OBJECTIVES: The aim of this study is to evaluate usability and reading accuracy of home pregnancy test formats. METHODS: Female volunteers, 18 - 45 years (Manchester, UK) completed questionnaires on their home-use experience of six pregnancy tests (strip, cassette, midstream visual and digital formats). These volunteers then evaluated device results using hCG-urine standards at a study centre, thereafter completing a questionnaire and ranking evaluation. RESULTS: Data were available from 111 volunteers. Women preferred midstream test formats; > 70% scored branded midstream digital and easy-use visual tests as 1or 2 (7-point Likert score), compared with ∼ 30% for store-brand and branded midstream visual tests, and < 10% for cassette or strip tests. Many cassette tests (23%) failed to provide a result (4, ≤ 2% for strips, midstream, respectively). Volunteers disagreed with study co-ordinator reading of test results in 30 and 40% of cases for the cassette and strip test results, respectively, compared with < 3% when using midstream digital or easy-use visual tests. Volunteers preferred the branded midstream digital, followed by branded midstream easy-use and visual tests. CONCLUSIONS: In this study, the branded midstream digital test was superior to other tests evaluated and fulfilled the criteria of being an easy-to-use and interpret test; strip and cassette tests showed poor performance in women's hands.

Comparison of a digital ovulation test with three popular line ovulation tests to investigate user accuracy and certainty.

OBJECTIVES: To determine the accuracy and certainty with which volunteers interpreted results of a digital ovulation test, Clearblue digital ovulation test (CB-DOT), compared with three home use non-digital visual ovulation tests: Clearblue ovulation test (CB-OT), First Response (FR) and Answer (AN). METHODS: A total of 72 female volunteers aged 18 - 45 years interpreted test results from each of the four ovulation tests to determine the day of the luteinising hormone surge in 40 individual menstrual cycles. We used urine previously collected from 25 volunteers. The accuracy with which volunteers interpreted the test results was calculated by comparing their results with results obtained by trained technicians using a blinded test regime. For each of the four tests, volunteers were also asked to rate seven attributes of certainty and eight attributes of preference. The primary objective was to compare the accuracy with which volunteers read results from CB-DOT when compared to three visual-based line ovulation tests. RESULTS: A significantly higher percentage of volunteers/technicians agreed on the interpretation of the results from CB-DOT (97.3%) than for CB-OT (83.5%; p = 0.0153), AN (73%; p = 0.0011) or FR (64.3%; p = 0.0001). CB-DOT was also found to have significantly better Likert scores than CB-OT, FR and AN for all seven attributes of certainty and was the test that 97.2% of volunteers preferred. CONCLUSIONS: Women can misread the results of line ovulation tests. Over 97% of volunteers correctly read the result of CB-DOT. CB-DOT was also the test that women read with most certainty and the test that most users preferred.